Novel compounds for diagnosis

Abstract

This invention relates to novel compounds of formula (I) or detectable labeled compounds thereof, stereoisomers, racemic mixtures, pharmaceutically acceptable salts, hydrates, or solvates, which can be used for imaging and determining the amount of α-synuclein aggregates. Furthermore, the compounds can be used to diagnose diseases, conditions, or abnormalities associated with α-synuclein aggregates (including, but not limited to, Lewy bodies and / or Lewy neurites) (e.g., Parkinson's disease), determine susceptibility to said diseases, conditions, or abnormalities, predict the prognosis of said diseases, conditions, or abnormalities, monitor disease progression in patients with said diseases, conditions, or abnormalities, monitor the progression of said diseases, conditions, or abnormalities, and predict the responsiveness of patients with said diseases, conditions, or abnormalities to their treatment.

Description

Invention Field

[0001] This invention relates to novel compounds of formula (I) or detectable labeled compounds thereof, stereoisomers, racemic mixtures, pharmaceutically acceptable salts, hydrates, or solvates, which can be used to image and determine the amount of α-synuclein aggregates. Furthermore, the compounds can be used to diagnose diseases, conditions, or abnormalities associated with α-synuclein (α-synuclein, A-synuclein, α-synuclein, A-syn, α-syn, aSyn, α-syn) aggregates, including but not limited to Lewy bodies and / or Lewy neurites (e.g., Parkinson's disease), determine susceptibility to said diseases, conditions, or abnormalities, predict the prognosis of said diseases, conditions, or abnormalities, monitor disease development in patients with said diseases, conditions, or abnormalities, monitor disease progression, and predict the responsiveness of patients with such diseases, conditions, or abnormalities to treatment. The invention also relates to methods for preparing said compounds and their precursors, diagnostic compositions comprising said compounds, methods for using said compounds, kits comprising said compounds, and uses thereof. Background of the Invention

[0003] Many age-related diseases are based on or associated with extracellular or intracellular deposits of amyloid material or amyloid protein, which contribute to disease pathogenesis and progression. The best-characterized amyloid protein for forming extracellular aggregates is β-amyloid (Abeta or Aβ).

[0004] Amyloid proteins that primarily form intracellular aggregates include, but are not limited to, Tau, α-synuclein, and huntingtin (HTT). Diseases involving α-synuclein aggregates are generally classified as synucleinopathy (or α-synucleinosis), and these include, but are not limited to, Parkinson's disease (PD). Synuclein diseases primarily involving neuronal aggregates include, but are not limited to, Parkinson's disease (sporadic, familial with SNCA mutations (the gene encoding α-synuclein) or SNCA gene duplication or triple duplication, familial with mutations in genes other than SNCA, simple autonomic failure, and Lewy body dysphagia), SNCA duplication carriers, Lewy body dementia (LBD), Lewy body dementia (DLB) (“simple” Lewy body dementia), Parkinson's disease dementia (PDD), diffuse Lewy body disease (DLBD), Alzheimer's disease, sporadic Alzheimer's disease, familial Alzheimer's disease with APP mutations, familial Alzheimer's disease with PS-1, PS-2, or other mutations, familial British dementia, Lewy body variants of Alzheimer's disease, and normal aging in Down syndrome. Synucleinopathy involving neurons and glial aggregates with α-synuclein includes, but is not limited to, multiple system atrophy (MSA) (Shy-Drager syndrome, striatal substantia nigra degeneration, and olivary pontocerebellar atrophy). Other diseases that may involve α-synuclein immune response disorders include, but are not limited to, traumatic brain injury, chronic injury encephalopathy, boxer's dementia, tau disorders (Pick disease, frontotemporal dementia, progressive supranuclear palsy, corticobasal degeneration and Niemann-Pick II disease type C1, frontotemporal dementia associated with chromosome 17 and Parkinson's syndrome), motor neuron disease, Huntington's disease, amyotrophic lateral sclerosis (sporadic, familial, and Guam ALS-dementia syndrome), axial dystrophy, and type 1 brain disorders. Iron-accumulating neurodegeneration (Hassler-Scheinker syndrome), prions, Creutzfeldt-Jakob disease, ataxia-telangiectasia, idiopathic orofacial motor disorder, subacute sclerosing panencephalitis, Gerstmann-Straussler-Scheinker syndrome, inclusion body myositis, Gaucher disease and other lysosomal storage disorders (including Kufor-Rakeb syndrome and Sanfilippo syndrome) and tachyophthalmos (REM) sleep behavior disorder (Jellinger, Mov Disord 2003, 18 Supplement 6, S2-12; Galvin et al. JAMA Neurology 2001, 58(2), 186-190; Kovari et al., Acta Neuropathol).2007, 114(3), 295-8; Saito et al., J Neuropathol Exp Neurol, 2004, 63(4), 323-328; McKee et al., Brain, 2013, 136(Pt 1), 43-64; Puschmann et al., Parkinsonism Relat Disord 2012, 18S1, S24-S27; Usenovic et al., J Neurosci. 2012, 32(12), 4240-4246; Winder-Rhodes et al., Mov Disord. 2012, 27(2), 312-315; Ferman et al., J Int Neuropsychol Soc. 2002, 8(7), 907-914; Smith et al., J Pathol. 2014; 232: 509-521; Lippa et al., Ann Neurol. Mar 1999; 45(3): 353-7; Schmitz et al., Mol Neurobiol. Aug 22, 2018; Charles et al., Neurosci Lett. July 28, 2000; 289(1): 29-32; Wilhelmsen et al., Arch Neurol. Mar 2004; 61(3): 398-406; Yamaguchi et al., J Neuropathol Exp Neurol. 2004, 80th Annual Meeting, Vol. 63; Askanas et al., J Neuropathol Exp Neurol. July 2000; 59(7): 592-8.

[0005] α-synuclein is a natural unfolded protein of 140 amino acids (Iwai et al., Biochemistry 1995, 34(32), 10139-10145). The sequence of α-synuclein can be divided into three main domains: 1) an N-terminal region containing residues 1-60, which contains 11-mer amphiphilic incomplete repeating residues (KTKEGV) with highly conserved hexamers. This region is involved in regulating the binding of α-synuclein to the membrane and its internalization; 2) a hydrophobic non-β amyloid component (NAC) domain spanning residues 61-95, which is crucial for α-synuclein fibrillation; and 3) a C-terminal region spanning residues 96-140, which is strongly acidic and proline-rich and has no obvious structural preference. α-synuclein has been shown to undergo multiple post-translational modifications, including truncation, phosphorylation, ubiquitination, oxidation, and / or transglutaminase covalent cross-linking (Fujiwara et al., Nat Cell Biol 2002, 4(2): 160-164; Hasegawa et al., J Biol Chem 2002, 277(50): 49071-49076; Li et al., Proc Natl Acad Sci US A 2005, 102(6): 2162-2167; Oueslati et al., ProgBrain Res 2010, 183: 115-145; Schmid et al., J Biol Chem 2009, 284(19): 13128-13142). Interestingly, most of these modifications involve residues in the C-terminal region.

[0006] Multiple phosphorylation sites have been detected in the C-terminal regions of Tyr-125, -133, and -136, as well as Ser-129 (Negro et al., FASEB J 2002, 16(2), 210-212). Tyr-125 residues can be phosphorylated by two Src family protein tyrosine kinases, c-Src and Fyn (Ellis et al., J Biol Chem 2001, 276(6), 3879-3884; Nakamura et al., Biochem Biophys Res Commun 2001, 280(4), 1085-1092). Phosphorylation of Src family kinases does not inhibit or enhance the tendency of α-synuclein polymerization. α-synuclein has been shown to be an in vitro protein tyrosine kinase p72. syk(Syk) is an outstanding matrix; once extensively phosphorylated by Syk or tyrosine kinases with similar specificity, it loses its ability to form oligomers, suggesting that these tyrosine kinases have a putative anti-neurodegenerative effect (Negro et al., FASEB J 2002, 16(2), 210-212). α-synuclein can be Ser-phosphorylated by protein kinases CKI and CKII (Okochi et al., J Biol Chem 2000, 275(1), 390-397). The residue Ser-129 can also be phosphorylated by G protein-coupled receptor protein kinases (Pronin et al., J Biol Chem 2000, 275(34), 26515-26522). In synucleopathic damage, including the Lewy body, extensive and selective phosphorylation of α-synuclein at Ser-129 is evident (Fujiwara et al., Nat Cell Biol 2002, 4(2); 160-164). Other post-translational modifications at the C-terminus, including glycosylation at Ser-129 (McLean et al., Neurosci Lett 2002, 323(3), 219-223) and nitration at Tyr-125, -133, and -136 (Takahashi et al., Brain Res 2002, 938(1-2), 73-80), can affect α-synuclein aggregation. It has been reported that truncation of the C-terminal region through proteolysis plays a role in the formation of α-synuclein protofibrils in various neurodegenerative diseases (Rochet et al., Biochemistry 2000, 39(35), 10619-10626). Full-length as well as partially truncated and insoluble α-synuclein aggregates have been detected in highly purified Lewy bodies (Crowther et al., FEBS Lett 1998, 436(3), 309-312).

[0007] Abnormal protein aggregation appears to be a common feature of the aging brain and a variety of neurodegenerative diseases (Trojanowski et al., 1998, Cell Death Differ. 1998, 5(10), 832-837; Koo et al., Proc Natl Acad Sci. 1999, 96(18), 9989-9990; Hu et al., Chin. Sci. Bull. 2001, 46, 1-3); although its exact role in the disease process remains to be determined. In in vitro models, α-synuclein (or some truncated forms thereof) readily assembles into filaments similar to those isolated from the brains of patients with Lewy body (LB) dementia and familial PD (Crowther et al., FEBS Lett 1998, 436(3), 309-312). α-synuclein and its mutant forms (A53T and A30P) have a random coil conformation and do not form significant secondary structures in low-concentration aqueous solutions; however, at higher concentrations, they tend to self-aggregate, producing amyloid fibrils (Wood et al., J Biol Chem 1999, 274(28), 19509-19512). Previous results have shown many differences in the aggregation behavior of PD-linked mutants and wild-type proteins. Monomeric α-synuclein aggregates in vitro in a metastable oligomeric state (i.e., fibrils) to form stable fibrils (Volles et al., Biochemistry 2002, 41(14), 4595-4602).

[0008] Parkinson's disease (PD) is the most common neurodegenerative motor disorder. PD is primarily an idiopathic disease, although in at least 5% of PD patients, the pathology is associated with mutations in one or more specific genes. Several point mutations in the α-synuclein gene (A30P, E46K, H50Q, G51D, A53T) have been described, causing familial PD with autosomal dominant inheritance. Furthermore, duplications and tripletuplets of the α-synuclein gene have been described in patients with PD, highlighting the role of α-synuclein in the pathogenesis of PD (Lesage et al., Hum. Mol. Genet., 2009, 18, R48-59). The pathogenesis of PD remains difficult to elucidate. However, mounting evidence suggests the role of pathogenic folding of α-synuclein, which leads to the formation of amyloid fibrils. In fact, the hallmarks of PD are primarily the presence of intracellular α-synuclein aggregates called Lewy bodies and neurites in substantia nigra neurons, and the death of dopaminergic neurons in the substantia nigra and other sites. α-synuclein is a naturally unfolded presynaptic protein that can misfold and aggregate into larger oligomers and fibrils associated with the pathogenesis of PD. Recent studies have suggested that small soluble oligomers and fibrils of α-synuclein are the most neurotoxic forms (Lashuel et al., J. Mol. Biol., 2002, 322, 1089-102). However, the exact role of α-synuclein in neuronal cytotoxicity remains to be elucidated (Review: Cookson, Annu. Rev. Biochem., 2005, 74, 29-52).

[0009] Besides Parkinson's disease, the accumulation of α-synuclein in the Lewy body is characteristic of all Lewy body disorders, including Parkinson's disease with dementia (PDD) and Lewy body dementia (DLB) (Capouch et al., Neurol Ther. 2018, 7, 249-263). In DLB, Lewy bodies are diffusely distributed throughout the cerebral cortex, and in addition to Lewy bodies and neurites, more linear and punctate structures (Lewy points) are found to be immunopositive for phosphorylated α-synuclein at Ser-129 (Outeiro et al., Mol Neurodegener. 2019, 14, 5). α-synuclein aggregates have also been found in multiple system atrophy (MSA), a rare and sporadic neurodegenerative disease characterized by rapidly progressive autonomic and motor dysfunction, as well as variable cognitive decline. These diseases include Shy-Drager syndrome, striatal substantia nigra degeneration, and olivary pontocerebellar atrophy. Depending on the dominant motor phenotype, this disease can be clinically reclassified as either Parkinson's disease (MSA-P) or cerebellar (MSA-C) variants (Fanciulli et al., N Engl J Med 2015; 372, 249-63). It is characterized by the accumulation of α-synuclein in the cytoplasm of oligodendrocytes, forming glial cytoplasmic inclusions (GCIs). GCIs, primarily composed of α-synuclein in fibrillary form, are a neuropathological marker of MSA and are found throughout the neocortex, hippocampus, brainstem, spinal cord, and dorsal root ganglia (Galvin et al., Arch Neurol. 2001, 58, 186-90). GCIs are considered central players in the pathogenesis of MSA. Correlation between GCI load and the degree of neuronal loss has been reported in the striatum, substantia nigra, and oligopontine-cerebellar regions (Stefanova et al., Neuropathol Appl Neurobiol. 2016, 42, 20-32). Furthermore, a causal relationship between GCI and the induction of neuronal loss has been demonstrated in transgenic mice overexpressing human α-synuclein in oligodendrocytes under various oligodendrocyte-specific promoters. A key event in the pathophysiological cascade is considered to be the permitted template formation of misfolded α-synuclein (“prion-like” transmission).

[0010] Diagnosis of Parkinson's disease is largely clinical and depends on the presence of a specific set of symptoms and signs (initial core features being bradykinesia, rigidity, resting tremor, and postural instability), the absence of atypical features, a slowly progressive course, and a response to symptomatic drug therapy, primarily limited to dopamine replacement therapy. Accurate diagnosis requires sophisticated clinical skills and is subject to a degree of subjectivity and error, as several other degenerative and non-degenerative diseases can mimic PD symptoms (multiple system atrophy (MSA), progressive supranuclear palsy (PSP), Alzheimer's disease (AD), essential tremor, dystonia tremor) (Guideline No. 113: Diagnosis and pharmacological management of Parkinson's disease, January 2010. SIGN). Final pathological confirmation can only be achieved through postmortem neuropathological analysis.

[0011] Brain scans on computed tomography (CT) and conventional magnetic resonance imaging (MRI) are usually normal in patients with Parkinson's disease (PD). However, these techniques can be used to rule out other conditions that may be secondary causes of PD, such as basal ganglia tumors, vascular lesions, and hydrocephalus. Specific MRI techniques (diffusion MRI) have been reported to be able to differentiate between typical and atypical PD, although their exact diagnostic value is still under investigation. Dopaminergic function in the basal ganglia can be measured using different PET and SPECT radiotracers. An example is iodofluoroane used in SPECT (…). 123 I) (trade name DaTSCAN) and iodothiophene (Dopascan) or fluorodeoxyglucose for PET ( 18 F)( 18 F-FDG) and dihydrobenzonazine ( 11 C)( 11 C-DTBZ). A pattern of reduced dopaminergic activity in the basal ganglia can aid in the diagnosis of PD, particularly in the symptomatic phase (Brooks, J. Nucl. Med., 2010, 51, 596-609; Redmond, Neuroscientist, 2002, 8, 457-88; Wood, Nat. Rev. Neurol., 2014, 10, 305).

[0012] Strategies are being developed to apply recent advances in understanding the underlying causes of Parkinson's disease to the development of biochemical biomarkers (Schapira Curr Opin Neurol 2013; 26(4): 395-400). Such biomarkers, which have been studied in various bodily fluids (cerebrospinal fluid (CSF), plasma, saliva), include α-synuclein levels, as well as DJ-1, τ, and Aβ, and neurofilaments, interleukins, osteopontin, and hypocrontin (Schapira Curr Opin Neurol 2013; 26(4): 395-400), but to date, none of these biomarkers, alone or in combination, are available for definitive diagnostic testing. To our knowledge, despite the urgent need for Parkinson's disease research and drug development, there are currently no approved α-synuclein diagnostic reagents available on the market or for clinical use (Eberling et al., J Parkinsons Dis. 2013; 3(4): 565-7).

[0013] The ability to image alpha-synuclein deposition in the brain would be a major achievement for alpha-synucleinopathy research, including Parkinson's disease research, diagnosis, and drug development. Accumulated alpha-synuclein in the brain is considered a key pathological marker of PD and can begin many years before symptoms appear. Therefore, alpha-synuclein is a priority target for drug development, not only considering its potential contribution to neurodegeneration but also because it offers the possibility of treating the disease while it remains asymptomatic or in its prodromal stages. In vivo imaging of alpha-synuclein pathology can be used as a biomarker to (i) potentially detect the presence of the disease at an early stage, (ii) assess disease progression, and (iii) serve as a pharmacodynamic tool for drug development. Today, considering the development of alpha-synuclein PET imaging agents, starting with the optimal selection of trial populations, is crucial for the accurate diagnosis of synucleinopathy and for supporting the clinical development of therapeutics targeting alpha-synuclein (Eberling, Dave and Frasier, J. Parkinson's Disease, 3, 565-567 (2013)). Despite significant efforts to identify α-synuclein PET ligands, only compounds that bind to synthetic α-synuclein fibrils with reasonably high affinity have been identified to date, but none of them have been validated in human clinical trials. They are not optimal for many reasons: low affinity or no binding is observed on pathological aggregates of α-synuclein present in diseased brains, low or no selectivity of α-synuclein relative to other aggregates has been reported, and their physicochemical properties are unsuitable for use as brain-penetrating PET reagents (Eberling et al., J Parkinsons Dis. 2013; 3(4): 565-7; Neal et al., Mol Imaging Biol. 2013; 15: 585-595; Bagchi et al., PLoS One 2013; 8(2): e55031; Yu et al., Bioorganic and Medicinal Chemistry 2012; 20: 4625-4634; Zhang et al., Appl Sci (Basel) 2014; 4(1): 66-78; Chu et al., J Med Chem, 2015, 58(15): 6002-17).

[0014] Therefore, it is clearly necessary to find molecular probes with high α-synuclein selectivity that recognize and bind to pathological α-synuclein. To reduce background signal interference caused by nonspecific off-target binding and to lower dosing requirements, α-synuclein imaging compounds should bind to their targets with high affinity and selectivity. For imaging α-synuclein aggregates associated with neurological diseases such as Parkinson's disease, imaging compounds need to penetrate the blood-brain barrier and enter the relevant brain regions. Cell permeability is another requirement for imaging compounds to target intracellular amyloid inclusions such as α-synuclein. Another prerequisite to avoid unnecessary accumulation of compounds that could increase the risk of unwanted side effects is rapid elution of the compound from the brain (or other target organs).

[0015] WO2011 / 128455 relates to specific compounds suitable for treating conditions associated with amyloid or amyloid-like proteins. US2012 / 0302755 relates to certain imaging agents for detecting neurological dysfunction. Other compounds for diagnosing neurodegenerative diseases of the olfactory epithelium are discussed in WO2012 / 037928.

[0016] WO2010 / 063701 relates to an in vivo imaging agent in a method for determining the presence or susceptibility to Parkinson's disease, wherein the in vivo imaging agent comprises an α-synuclein binding agent labeled with an in vivo imaging component, wherein the in vivo imaging agent binds to α-synuclein with binding affinity.

[0017] US2014 / 0142089 relates to methods for preventing or treating degenerative brain diseases, the method comprising administering an effective amount of a pharmaceutical composition to an individual in need, said pharmaceutical composition comprising a particular compound, a pharmaceutically acceptable salt thereof, an isomer thereof, a solvate thereof, a hydrate thereof, or a combination thereof.

[0018] WO 2009 / 155017 describes aryl or heteroaryl substituted azirzooxazole derivatives, which are claimed to be used as tracers in positron emission tomography (PET) imaging for studying amyloid deposits in the brain in vivo for the diagnosis of Alzheimer's disease.

[0019] WO 2016 / 033445 relates to specific compounds used for imaging the huntingtin protein.

[0020] WO2017 / 153601 and WO 2019 / 234243 relate to bicyclic compounds for the diagnosis of α-synuclein aggregates.

[0021] Surprisingly, a new class of compounds of formula (I) or its subforms (e.g., (IIa), (IIb), (IIIa), (IIIb), (IIIc), (III-F), (III-H)) or their detectably labeled compounds, stereoisomers, racemic mixtures, pharmaceutically acceptable salts, hydrates, or solvates are found to bind α-synuclein. Therefore, when radiolabeled with a suitable radioisotope, the compounds of this invention are qualified as PET tracers for imaging pathological α-syn aggregates in PD and other α-synucleinopathies. Invention Overview

[0023] One object of the present invention is to provide compounds that can be used to diagnose diseases, conditions, or abnormalities associated with α-synuclein aggregates, including but not limited to Lewy bodies and / or Lewy neurites (e.g., Parkinson's disease), predict the prognosis of said diseases, conditions, or abnormalities, and monitor the progression of said diseases, conditions, or abnormalities. In particular, said compounds should be suitable for determining susceptibility to said diseases, conditions, or abnormalities, monitoring the progression of said diseases, conditions, or abnormalities, or predicting the response of patients with said diseases, conditions, or abnormalities to treatment with a certain drug.

[0024] Furthermore, there is a need for compounds that can be used as imaging agents for α-synuclein aggregates (including but not limited to Lewy bodies and / or Lewy neurites). In particular, an object of the present invention is to provide compounds suitable for use in diagnostic compositions for positron emission tomography imaging of α-synuclein diseases, for example, wherein... 18 The compound is detectably labeled with F or other markings.

[0025] The inventors have surprisingly discovered that these objectives can be achieved by compounds of formula (I) or its subforms (e.g., (IIa), (IIb), (IIIa), (IIIb), (IIIc), (III-F), (III-H)) as described below, or by compounds of the same or detectably labeled form, stereoisomers, racemic mixtures, pharmaceutically acceptable salts, hydrates or solvates.

[0026] Compounds of formula (I) or its subforms (e.g., (IIa), (IIb), (IIIa), (IIIb), (IIIc), (III-F), (III-H)) or their detectable labeled compounds, stereoisomers, racemic mixtures, pharmaceutically acceptable salts, hydrates, or solvates exhibit potent binding affinity for α-synuclein aggregates in mammalian (e.g., human) tissues. Furthermore, compounds of formula (I) or its subforms (e.g., (IIa), (IIb), (IIIa), (IIIb), (IIIc), (III-F), (III-H)) or their detectable labeled compounds, stereoisomers, racemic mixtures, pharmaceutically acceptable salts, hydrates, or solvates exhibit potent selectivity for α-synuclein relative to other protein aggregates associated with neurodegeneration, enabling differentiation of PD from other protein diseases sharing common clinical and pathological features. Due to their unique design features, these compounds exhibit properties such as appropriate lipophilicity and molecular weight, brain uptake and pharmacokinetics, cell permeability, solubility and autofluorescence, making them successful imaging probes for the in vivo, in vitro and ex vivo detection and quantification of α-synuclein aggregates (including but not limited to Lewy bodies and / or Lewy neurites).

[0027] This invention discloses novel compounds of formula (I) or its subforms (e.g., (IIa), (IIb), (IIIa), (IIIb), (IIIc), (III-F), (III-H)) disclosed herein, or detectable labeled compounds, stereoisomers, racemic mixtures, pharmaceutically acceptable salts, hydrates, or solvates thereof, which exhibit enhanced binding properties to α-synuclein aggregates (including, but not limited to, Lewy bodies and / or Lewy neurites). The compounds of this invention can be labeled (e.g., radiolabeled) so that they can be used for in vitro, ex vivo, and in vivo imaging to detect α-synuclein aggregates, including, but not limited to, Lewy bodies and / or Lewy neurites. This invention provides a method for detecting α-synuclein aggregates, including but not limited to Lewy bodies and / or Lewy neurites, in vitro using compounds of formula (I) or its subforms (e.g., (IIa), (IIb), (IIIa), (IIIb), (IIIc), (III-F), (III-H)) or their detectable labeled compounds, stereoisomers, racemic mixtures, pharmaceutically acceptable salts, hydrates, or solvates, or pharmaceutical compositions thereof. This invention also provides a method for using compounds of formula (I) or its subforms (e.g., (IIa), (IIb), (IIIa), (IIIb), (IIIc), (III-F), (III-H)) or their detectable labeled compounds, stereoisomers, racemic mixtures, pharmaceutically acceptable salts, hydrates, or solvates, as diagnostic imaging agents, particularly for detecting pre-symptom or prodromal symptoms of Parkinson's disease and / or other synucleinogenic disorders, for example using positron emission tomography (PET). The compounds of the present invention can be used as biomarkers for monitoring local anatomy and transient progression of pathological conditions, leading to improvements in the design and outcomes of clinical diagnostic studies. The present invention also provides diagnostic compositions comprising compounds of formula (I) or its subforms (e.g., (IIa), (IIb), (IIIa), (IIIb), (IIIc), (III-F), (III-H)) or detectable labeled compounds thereof, stereoisomers, racemic mixtures, pharmaceutically acceptable salts, hydrates or solvates, and at least one pharmaceutically acceptable excipient, carrier, diluent, or adjuvant.

[0028] The present invention is summarized in the following scheme:

[0029] This invention relates to compounds of formula (I):

[0030]

[0031] Or its detectable labeled compounds, stereoisomers, racemic mixtures, pharmaceutically acceptable salts, hydrates, or solvates, wherein

[0032] It is an aryl or heteroaryl group, oriented and selected from the following:

[0033]

[0034] R 0 It is H or C1-C4 alkyl;

[0035] R 1 -CN; or halogen; or C1-C4 alkyl; or C1-C4 alkoxy; or -N(C1-C4 alkyl)2; or -NH(C1-C4 alkyl); or H; or

[0036] R 1 It is a -NH-C3-C6 cycloalkyl, C3-C6 cycloalkyl or heterocyclic group, each of which is optionally substituted with at least one halogen;

[0037] R 2 It is an aryl, 5-membered, or 6-membered heteroaryl group, wherein R 2 Selected from the following:

[0038]

[0039] in

[0040] R 2a R 2a’ Independently selected from H or F;

[0041] R 2b Independently selected from F, -OH, C1-C4 alkyl, halo-C1-C4 alkyl, -NH2, -CN or C1-C4 alkoxy;

[0042] R 2c R 2c’ It is independently selected from H, F, OH, OCH3 or CH3;

[0043] R 2d Selected from H, F, or -OH;

[0044] R 2e Selected from H, OH, CH3 or F;

[0045] Z can be N, NH, N (C1-C4 alkyl), N (halogenated C1-C4 alkyl), O, or S independently;

[0046] Z 1 It can be N, NH, O, or S independently;

[0047] p is 0, 1, or 2;

[0048] m is 0 or 1;

[0049] When valence is allowed It is a combination of single and double bonds; and

[0050] * indicates the location of the bond.

[0051] In another aspect, the present invention also relates to compounds having the following formula

[0052]

[0053] Or its detectable labeled compounds, stereoisomers, racemic mixtures, pharmaceutically acceptable salts, hydrates, or solvates.

[0054] In another aspect, the present invention also relates to compounds having the following formula

[0055] Or its detectable labeled compounds, stereoisomers, racemic mixtures, pharmaceutically acceptable salts, hydrates, or solvates.

[0056] In one aspect, a compound of formula (I) or its sub-formulas (e.g., (IIa), (IIb), (IIIa), (IIIb), (IIIc), (III-F), (III-H)) or a detectable labeled compound thereof, stereoisomer, racemic mixture, pharmaceutically acceptable salt, hydrate or solvate is used for imaging α-synuclein aggregates (including but not limited to Lewy bodies and / or Lewy neurites), wherein said compound is preferably used for positron emission tomography imaging of α-synuclein aggregates (including but not limited to Lewy bodies and / or Lewy neurites).

[0057] In another aspect, the present invention relates to a method for imaging diseases, conditions, or abnormalities associated with α-synuclein aggregates in an individual, said α-synuclein aggregates including, but not limited to, Lewy bodies and / or Lewy neurites, the method comprising the following steps:

[0058] (a) administering to an individual a compound of formula (I) or its subforms (e.g. (IIa), (IIb), (IIIa), (IIIb), (IIIc), (III-F), (III-H)) or a detectable labeled compound thereof, stereoisomer, racemic mixture, pharmaceutically acceptable salt, hydrate or solvate;

[0059] (b) to bind the compound to α-synuclein aggregates (including but not limited to Lewy bodies and / or Lewy neurites); and

[0060] (c) Detecting compounds that bind to α-synuclein aggregates (including but not limited to Lewy bodies and / or Lewy neurites).

[0061] In another aspect, the present invention relates to a method for imaging diseases, conditions, or abnormalities associated with α-synuclein aggregates in an individual, said α-synuclein aggregates including, but not limited to, Lewy bodies and / or Lewy neurites, the method comprising the following steps:

[0062] (a) administering to an individual a compound of formula (I) or its subforms (e.g., (IIa), (IIb), (IIIa), (IIIb), (IIIc), (III-F), (III-H)) or a detectably labeled compound thereof, stereoisomer, racemic mixture, pharmaceutically acceptable salt, hydrate, or solvate; and

[0063] (b) To image the brain of the individual.

[0064] In another aspect, the present invention relates to a method for positron emission tomography (PET) imaging of α-synuclein aggregates (including but not limited to Lewy bodies and / or Lewy neurites) in individual tissues, the method comprising the following steps:

[0065] (a) administering to an individual a compound of formula (I) or its subforms (e.g. (IIa), (IIb), (IIIa), (IIIb), (IIIc), (III-F), (III-H)) or a detectable labeled compound thereof, stereoisomer, racemic mixture, pharmaceutically acceptable salt, hydrate or solvate;

[0066] (b) to allow the compound to penetrate into the tissues of the individual; and

[0067] (c) Acquire positron emission tomography (PET) images of the tissue of the individual; wherein the tissue is tissue of the central nervous system (vNS), tissue of the eye or brain, preferably wherein the tissue is brain tissue.

[0068] In another aspect, the present invention relates to a method for detecting neurological diseases, conditions, or abnormalities associated with α-synuclein aggregates in an individual, said α-synuclein aggregates including, but not limited to, Lewy bodies and / or Lewy neurites, the method comprising the following steps:

[0069] (a) administering to an individual a compound of formula (I) or its subforms (e.g. (IIa), (IIb), (IIIa), (IIIb), (IIIc), (III-F), (III-H)) or a detectable labeled compound thereof, stereoisomer, racemic mixture, pharmaceutically acceptable salt, hydrate or solvate;

[0070] (b) to bind the compound to α-synuclein aggregates (including but not limited to Lewy bodies and / or Lewy neurites); and

[0071] (c) Measure the radioactive signal of the compound, which binds to the α-synuclein aggregate (including but not limited to Lewy bodies and / or Lewy neurites).

[0072] In another aspect, the present invention relates to a method for detecting and / or quantifying α-synuclein aggregates in individual tissues, said α-synuclein aggregates including, but not limited to, Lewy bodies and / or Lewy neurites, the method comprising the following steps:

[0073] (a) Contact the individual tissue with a compound of formula (I) or its subforms (e.g., (IIa), (IIb), (IIIa), (IIIb), (IIIc), (III-F), (III-H)) or a detectably labeled compound thereof, stereoisomer, racemic mixture, pharmaceutically acceptable salt, hydrate or solvate;

[0074] (b) to bind the compound to α-synuclein aggregates (including but not limited to Lewy bodies and / or Lewy neurites); and

[0075] (c) Using positron emission tomography to detect and / or quantify compounds that bind to α-synuclein aggregates, including but not limited to Lewy bodies and / or Lewy neurites.

[0076] In another aspect, the present invention relates to a method for diagnostic imaging of an individual's brain, the method comprising the following steps:

[0077] (a) administering to an individual a compound of formula (I) or its subforms (e.g., (IIa), (IIb), (IIIa), (IIIb), (IIIc), (III-F), (III-H)) or a detectably labeled compound thereof, stereoisomer, racemic mixture, pharmaceutically acceptable salt, hydrate, or solvate; and

[0078] (b) Images of the individual’s brain were obtained using positron emission tomography.

[0079] This invention also relates to a method for collecting data for diagnosing diseases, conditions, or abnormalities associated with α-synuclein aggregates (including but not limited to Lewy bodies and / or Lewy neurites), which is also disclosed herein, wherein the method comprises the following steps:

[0080] (a) Contact a sample or specific body part or region suspected of containing α-synuclein aggregates (including but not limited to Lewy bodies and / or Lewy neurites) with a compound of formula (I) or its subforms (e.g. (IIa), (IIb), (IIIa), (IIIb), (IIIc), (III-F), (III-H)) or its detectable labeled compounds, stereoisomers, racemic mixtures, pharmaceutically acceptable salts, hydrates or solvates;

[0081] (b) To bind the compound to α-synuclein aggregates (including but not limited to Lewy bodies and / or Lewy neurites);

[0082] (c) Detecting compounds that bind to α-synuclein aggregates (including but not limited to Lewy bodies and / or Lewy neurites); and

[0083] (d) Optionally establish a correlation between the presence or absence of a compound that binds to α-synuclein aggregates (including but not limited to Lewy bodies and / or Lewy neurites) and the presence or absence of α-synuclein aggregates (including but not limited to Lewy bodies and / or Lewy neurites) in a sample or a specific body part or region.

[0084] This invention also relates to a method for collecting data to determine susceptibility to diseases, conditions, or abnormalities associated with α-synuclein aggregates, including but not limited to Lewy bodies and / or Lewy neurites, the method comprising the following steps:

[0085] (a) Contact a sample or specific body part or region suspected of containing α-synuclein aggregates (including but not limited to Lewy bodies and / or Lewy neurites) with a compound of formula (I) or its subforms (e.g. (IIa), (IIb), (IIIa), (IIIb), (IIIc), (III-F), (III-H)) or its detectable labeled compounds, stereoisomers, racemic mixtures, pharmaceutically acceptable salts, hydrates or solvates;

[0086] (b) To bind the compound to α-synuclein aggregates (including but not limited to Lewy bodies and / or Lewy neurites);

[0087] (c) Detecting compounds that bind to α-synuclein aggregates (including but not limited to Lewy bodies and / or Lewy neurites); and

[0088] (d) Optionally establish a correlation between the presence or absence of a compound that binds to α-synuclein aggregates (including but not limited to Lewy bodies and / or Lewy neurites) and the presence or absence of α-synuclein aggregates (including but not limited to Lewy bodies and / or Lewy neurites) in a sample or a specific body part or region.

[0089] In another aspect, the present invention relates to a method for acquiring data for predicting the prognosis of diseases, conditions, or abnormalities associated with α-synuclein aggregates, including but not limited to Lewy bodies and / or Lewy neurites, wherein the method comprises the following steps:

[0090] (a) Contacting a sample, specific body part or region suspected of containing α-synuclein aggregates (including but not limited to Lewy bodies and / or Lewy neurites) with a compound of formula (I) or its subforms (e.g. (IIa), (IIb), (IIIa), (IIIb), (IIIc), (III-F), (III-H)) or its detectable labeled compounds, stereoisomers, racemic mixtures, pharmaceutically acceptable salts, hydrates or solvates;

[0091] (b) To bind the compound to α-synuclein aggregates (including but not limited to Lewy bodies and / or Lewy neurites);

[0092] (c) Detecting compounds that bind to α-synuclein aggregates (including but not limited to Lewy bodies and / or Lewy neurites);

[0093] (d) Optionally, establish a correlation between the presence or absence of compounds that bind to α-synuclein aggregates (including but not limited to Lewy bodies and / or Lewy neurites) and the presence or absence of α-synuclein aggregates (including but not limited to Lewy bodies and / or Lewy neurites) in the sample or a specific body part or region; and

[0094] (e) Optionally repeat steps (a) through (c) and optional step (d) (if any) at least once.

[0095] In another aspect, the present invention relates to a method for collecting data for monitoring the progression of a disease, condition, or abnormality associated with α-synuclein aggregates in patients, said α-synuclein aggregates including, but not limited to, Lewy bodies and / or Lewy neurites, the method comprising the following steps:

[0096] (a) Contacting a sample, specific body part or region suspected of containing α-synuclein aggregates (including but not limited to Lewy bodies and / or Lewy neurites) with a compound of formula (I) or its subforms (e.g. (IIa), (IIb), (IIIa), (IIIb), (IIIc), (III-F), (III-H)) or its detectable labeled compounds, stereoisomers, racemic mixtures, pharmaceutically acceptable salts, hydrates or solvates;

[0097] (b) To bind the compound to α-synuclein aggregates (including but not limited to Lewy bodies and / or Lewy neurites);

[0098] (c) Detecting compounds that bind to α-synuclein aggregates (including but not limited to Lewy bodies and / or Lewy neurites);

[0099] (d) Optionally, establish a correlation between the presence or absence of compounds that bind to α-synuclein aggregates (including but not limited to Lewy bodies and / or Lewy neurites) and the presence or absence of α-synuclein aggregates (including but not limited to Lewy bodies and / or Lewy neurites) in the sample or a specific body part or region; and

[0100] (e) Optionally repeat steps (a) through (c) and optional step (d) (if any) at least once.

[0101] In another aspect, the present invention relates to a method for collecting data to predict the responsiveness of a patient suffering from a disease, condition, or abnormality associated with α-synuclein aggregates (including but not limited to Lewy bodies and / or Lewy neurites) to treatments for a disease, condition, or abnormality associated with α-synuclein aggregates (including but not limited to Lewy bodies and / or Lewy neurites), wherein the α-synuclein aggregates include, but are not limited to, Lewy bodies and / or Lewy neurites, the method comprising the following steps:

[0102] (a) Contacting a sample, specific body part or region suspected of containing α-synuclein aggregates (including but not limited to Lewy bodies and / or Lewy neurites) with a compound of formula (I) or its subforms (e.g. (IIa), (IIb), (IIIa), (IIIb), (IIIc), (III-F), (III-H)) or its detectable labeled compounds, stereoisomers, racemic mixtures, pharmaceutically acceptable salts, hydrates or solvates;

[0103] (b) To bind the compound to α-synuclein aggregates (including but not limited to Lewy bodies and / or Lewy neurites);

[0104] (c) Detecting compounds that bind to α-synuclein aggregates (including but not limited to Lewy bodies and / or Lewy neurites);

[0105] (d) Optionally, establish a correlation between the presence or absence of compounds that bind to α-synuclein aggregates (including but not limited to Lewy bodies and / or Lewy neurites) and the presence or absence of α-synuclein aggregates (including but not limited to Lewy bodies and / or Lewy neurites) in the sample or a specific body part or region; and

[0106] (e) Optionally repeat steps (a) through (c) and optional step (d) (if any) at least once.

[0107] The present invention also relates to diagnostic or pharmaceutical compositions comprising compounds of formula (I) or its subforms (e.g., (IIa), (IIb), (IIIa), (IIIb), (IIIc), (III-F), (III-H)) or detectable labeled compounds thereof, stereoisomers, racemic mixtures, pharmaceutically acceptable salts, hydrates or solvates, and at least one pharmaceutically acceptable excipient, carrier, diluent or adjuvant.

[0108] In another aspect, the present invention also relates to compounds of formula (IV-F).

[0109]

[0110] Or its detectable labeled compounds, stereoisomers, racemic mixtures, pharmaceutically acceptable salts, hydrates, or solvates, wherein

[0111] R 3 Selected from

[0112] R 4 It is an aryl, 5-membered, or 6-membered heteroaryl group, wherein R 4 Selected from

[0113]

[0114] in

[0115] R 2a R 2a’ Independently selected from H or F;

[0116] R 2b Independently selected from F, -OH, C1-C4 alkyl, halo-C1-C4 alkyl, -NH2, -CN or C1-C4 alkoxy;

[0117] R 2c R 2c’It is independently selected from H, F, OH, OCH3 or CH3;

[0118] R 2d Selected from H, F, or -OH;

[0119] R 2e Selected from H, OH, CH3 or F;

[0120] Z can be N, NH, N (C1-C4 alkyl), N (halogenated C1-C4 alkyl), O, or S independently;

[0121] Z 1 It can be N, NH, O, or S independently;

[0122] p is 0, 1, or 2;

[0123] m is 0 or 1;

[0124] When valence is allowed For combinations of single and double bonds; and

[0125] * indicates the location of the bond.

[0126] In another aspect, the present invention also relates to compounds of formula (IV-H).

[0127]

[0128] Or its detectable labeled compounds, stereoisomers, racemic mixtures, pharmaceutically acceptable salts, hydrates, or solvates, wherein

[0129] R 5 Selected from

[0130] R 6 It is an aryl or 5- or 6-membered heteroaryl, wherein R 6 Selected from the following:

[0131]

[0132] in

[0133] R 2a R 2a’ Independently selected from H, X, or F;

[0134] R 2b Independently selected from X, F, -OH, C1-C4 alkyl, halo-C1-C4 alkyl, -NH2, -CN or C1-C4 alkoxy, wherein the C1-C4 alkyl, halo-C1-C4 alkyl or C1-C4 alkoxy optionally includes one or more X;

[0135] R2c R 2c’ It is independently selected from X, H, F, OH, OCH3 or CH3;

[0136] R 2d Selected from X, H, F, or -OH;

[0137] R 2e Selected from X, H, OH, CH3, or F;

[0138] Z can be N, NH, N (C1-C4 alkyl), N (halogenated C1-C4 alkyl), O, or S independently;

[0139] Z 1 It can be N, NH, O, or S independently;

[0140] p is 0, 1, or 2;

[0141] m is 0 or 1;

[0142] When valence is allowed It is a combination of single and double bonds;

[0143] * indicates the location of the bond;

[0144] Fluorine is 19 F;

[0145] X is bromine, chlorine, or iodine; and

[0146] Where R 6 It contains at least one X.

[0147] In another aspect, the present invention also relates to a method for preparing compounds of formula (III-F) or detectable labeled compounds thereof, stereoisomers, racemic mixtures, pharmaceutically acceptable salts, hydrates, or solvates, the method comprising reacting a compound of formula (IV-F) with... 18 The F-fluorinating agent reaction causes the leaving group (LG) to be... 18 F substitution.

[0148] This invention also relates to a method for preparing compounds of formula (III-H) or detectable labeled compounds thereof, stereoisomers, racemic mixtures, pharmaceutically acceptable salts, hydrates, or solvates, the method comprising reacting a compound of formula (IV-H) with a tritizing agent such that X is... 3 H substitution.

[0149] In another aspect, the present invention also relates to compounds of formula (IV-J),

[0150]

[0151] Or its stereoisomers, racemic mixtures, pharmaceutically acceptable salts, hydrates or solvates, wherein

[0152] R 7 Selected from

[0153] R 8 Selected from the following:

[0154]

[0155] in

[0156] R 2a R 2a’ Independently selected from H or F;

[0157] R 2b Independently selected from F, -OH, C1-C4 alkyl, halo-C1-C4 alkyl, -NH2, -CN or C1-C4 alkoxy;

[0158] p is 0, 1, or 2;

[0159] R z Selected from H, C1-C4 alkyl, or halo-C1-C4 alkyl;

[0160] When valence is allowed It is a combination of single and double bonds;

[0161] Fluorine is 19 F; and

[0162] * indicates the location of the bond.

[0163] In another aspect, the present invention also relates to a method for preparing compounds of formula (III-H) or detectable labeled compounds thereof, stereoisomers, racemic mixtures, pharmaceutically acceptable salts, hydrates, or solvates, comprising mixing a compound of formula (IV-J) with... 3 H radiolabeled reagent reaction.

[0164] The present invention also relates to the use of compounds of formula (I) or its subforms (e.g., (IIa), (IIb), (IIIa), (IIIb), (IIIc), (III-F), (III-H)) or their detectable labeled compounds, stereoisomers, racemic mixtures, pharmaceutically acceptable salts, hydrates or solvates as in vitro analytical references or in vitro screening tools.

[0165] The present invention also relates to a test kit for detecting and / or diagnosing diseases, conditions or abnormalities associated with α-synuclein aggregates, wherein the test kit comprises at least one compound of formula (I) or its subforms (e.g., (IIa), (IIb), (IIIa), (IIIb), (IIIc), (III-F), (III-H)) or a detectable labeled compound thereof, stereoisomer, racemic mixture, pharmaceutically acceptable salt, hydrate or solvate.

[0166] The present invention also relates to a kit for preparing radiopharmaceutical formulations, wherein the kit comprises a sealed vial containing at least one compound of formula (IV-F) or (IV-H) or (IV-J) or a compound thereof that is detectably labeled therewith, a stereoisomer, a racemic mixture, a pharmaceutically acceptable salt, a hydrate, or a solvate.

[0167] In the following text, compounds of formula (I) or its subforms (e.g., (IIa), (IIb), (IIIa), (IIIb), (IIIc), (III-F), (III-H)) or their detectably labeled compounds, stereoisomers, racemic mixtures, pharmaceutically acceptable salts, hydrates, or solvates are referred to as compounds of the present invention. Compounds of formulas (IV-F), (IV-H), and (IV-J) are referred to as precursors of compounds of the present invention.

[0168] The present invention is also defined by the following schemes:

[0169] A1. Compounds of formula (I)

[0170]

[0171] And all detectable labeled compounds, stereoisomers, racemic mixtures, pharmaceutically acceptable salts, hydrates, or solvates,

[0172] in

[0173] R 1 The following are fluorinated pyrrolidines

[0174] R 2 It is a 5- or 6-membered heteroaryl group containing one or two N atoms, wherein the heteroaryl group is optionally substituted with a methyl group, and

[0175] * indicates the location of the bond.

[0176] A2. Compounds of formula (I) of scheme A1, wherein

[0177] R 1 For the following 18 F-substituted pyrrolidines

[0178] A3. Compounds of formula (I) of scheme A1, wherein

[0179] R 1 For the following 19 F-substituted pyrrolidines and

[0180] The compound of formula (I) is at least one available position 3 H (tritium) can detect ground markers.

[0181] A4. The compound of any one of schemes A1-A3 is...

[0182]

[0183] A5. A compound of any one of schemes A1-A4, used for imaging α-synuclein aggregates (including but not limited to Lewy bodies and / or Lewy neurites), wherein the compound is preferably used for positron emission tomography imaging of α-synuclein aggregates, including but not limited to Lewy bodies and / or Lewy neurites.

[0184] A6. A compound of any one of schemes A1-A4, for the diagnosis of diseases, conditions, or abnormalities or susceptibility to α-synuclein aggregates (including, but not limited to, Lewy bodies and / or Lewy neurites), wherein said condition is optionally selected from Parkinson's disease (including sporadic, familial with α-synuclein mutations, familial with mutations other than α-synuclein, isolated autonomic dysfunction, or Lewy body dysphagia), Lewy body dementia (LBD), Lewy body dementia (DLB) (including “isolated” Lewy body dementia), Parkinson's disease dementia (PDD), diffuse Lewy body disease (DLBD), sporadic Alzheimer's disease, familial Alzheimer's disease with APP mutations, familial Alzheimer's disease with PS-1, PS-2, or other mutations, familial British dementia, Lewy body variants of Alzheimer's disease, Down syndrome, and multiple system atrophy (including Shy-Drager syndrome, striatum-nigrostriatum degeneration, or olivary-pontocerebellar atrophy). Traumatic brain injury, chronic traumatic encephalopathy, boxer's dementia, tau protein diseases (including Pick's disease, frontotemporal dementia, progressive supranuclear palsy, corticobasal degeneration, Niemann-Pick type C1 disease, frontotemporal dementia associated with Parkinson's syndrome related to chromosome 17), Creutzfeldt-Jakob disease, Huntington's disease, motor neuron disease, amyotrophic lateral sclerosis (including sporadic, familial, or Guam ALS-dementia syndrome), neural axis dystrophy, and neurological disorders with type I brain iron accumulation. Degenerative diseases (including Haas-Schwarz syndrome), prions, ataxia-telangiectasia, idiopathic orofacial motor disorders, subacute sclerosing panencephalitis, Gerstmann-Straussler-Scheatz syndrome, inclusion body myositis, Gaucher disease, Krab disease and other lysosomal storage diseases (including Kufor-Rakeb syndrome and Sanfilippo syndrome) and kinesiology (REM) sleep behavior disorders, with Parkinson's disease being the preferred option.

[0185] A7. A method for collecting data on diseases, conditions, or abnormalities associated with α-synuclein aggregates in a sample or patient for diagnostic purposes, wherein the α-synuclein aggregates include, but are not limited to, Lewy bodies and / or Lewy neurites, the method comprising:

[0186] (a) Contact a sample or specific body part or region suspected of containing α-synuclein aggregates (including but not limited to Lewy bodies and / or Lewy neurites) with a compound as defined in any of schemes A1-A4;

[0187] (b) To bind the compound to α-synuclein aggregates (including but not limited to Lewy bodies and / or Lewy neurites);

[0188] (c) Detecting compounds that bind to α-synuclein aggregates, said α-synuclein aggregates comprising Lewy bodies and / or Lewy neurites; and

[0189] (d) Optionally establish a correlation between the presence or absence of compounds that bind to α-synuclein aggregates (including but not limited to Lewy bodies and / or Lewy neurites) and the presence or absence of α-synuclein aggregates (including but not limited to Lewy bodies and / or Lewy neurites).

[0190] A8. A diagnostic composition comprising a compound of any one of schemes A1-A4 and a pharmaceutically acceptable excipient, carrier, diluent or adjuvant.

[0191] A9. Compounds of formula (II-F)

[0192]

[0193] And all detectable labeled compounds, stereoisomers, racemic mixtures, pharmaceutically acceptable salts, hydrates, or solvates,

[0194] in

[0195] R 3 The following are pyrrolidines substituted with leaving groups (LG).

[0196] R 4 It is a 5- or 6-membered heteroaryl group containing one or two N atoms, wherein the heteroaryl group is optionally substituted with a methyl group, and

[0197] * indicates the location of the bond.

[0198] A10. Compounds of formula (II-F) of scheme A9, wherein LG is selected from halogens, C 1-4 Alkyl sulfonates and C 6-10 Aryl sulfonates.

[0199] A11. Compounds of formula (II-F) of schemes A9 or A10, which are

[0200]

[0201] A12. Compounds of formula (II-H)

[0202]

[0203] And all detectable labeled compounds, stereoisomers, racemic mixtures, pharmaceutically acceptable salts, hydrates, or solvates,

[0204] in

[0205] R 5 The following are fluorinated pyrrolidines

[0206] R 6 It is a 5- or 6-membered heteroaryl group containing one or two N atoms, wherein the heteroaryl group is optionally substituted with a methyl group and / or the heteroaryl group is optionally substituted with one or more X atoms.

[0207] X is a halogen or H, provided that at least one X is a halogen, and

[0208] * indicates the location of the bond.

[0209] A13. The compound of formula (II-H) of scheme A12, which is

[0210]

[0211] A14. A method for preparing the compound of scheme A2, comprising mixing the compound of any one of schemes A9-A11 with... 18 The F-fluorinating agent reaction causes LG to be... 18 F substitution.

[0212] A15. The method of scheme 14, in which... 18 F-fluorinating agents are selected from K 18 F, H 18 F, Cs 18 F, Na 18 F and [ 18 F]Tetrabutylammonium fluoride.

[0213] A16. Use of any compound from schemes A1-A4 as an in vitro analytical reference or in vitro screening tool.

[0214] A17. A test kit for detecting and / or diagnosing conditions or abnormalities associated with α-synuclein aggregates, wherein the test kit contains at least one compound as defined in any one of schemes A1-A4.

[0215] A18. A kit for preparing radiopharmaceutical formulations, wherein the kit comprises a sealed vial containing at least one compound as defined in any one of schemes A9-A11.

[0216] The present invention is also defined by the following schemes:

[0217] B1. Compounds of formula (I)

[0218]

[0219] Or its detectable labeled compounds, stereoisomers, racemic mixtures, pharmaceutically acceptable salts, hydrates, or solvates.

[0220] in

[0221] R 1 The following are fluorinated pyrrolidines

[0222] R 2 It is a 5- or 6-membered heteroaryl group containing one or two N atoms, wherein the heteroaryl group is optionally substituted with a methyl group, and

[0223] * indicates the location of the bond.

[0224] B2. A compound of formula (I) of scheme B1, wherein the compound is a detectable labeled compound.

[0225] B3. A compound of formula (I) of scheme B2, wherein the detectable labeling compound comprises a detectable label selected from radioactive isotopes, preferably 2 H, 3 H or 18 F.

[0226] B4. Compounds of formula (I) of scheme B3, wherein

[0227] R 1 For the following 18 F-substituted pyrrolidines

[0228] B5. Compounds of formula (I) of scheme B3, wherein

[0229] R 1 For the following 19 F-substituted pyrrolidines and

[0230] The compound of formula (I) is at least one available position. 3 H (tritium) can detect ground markers.

[0231] B6. A compound from any one of schemes B1-B5, which is

[0232]

[0233] Where T refers to 3 H (tritium), and F refers to 19 F.

[0234] B7. A compound of any one of schemes B1-B6, used for imaging α-synuclein aggregates (including but not limited to Lewy bodies and / or Lewy neurites), wherein the compound is preferably used for positron emission tomography imaging of α-synuclein aggregates (including but not limited to Lewy bodies and / or Lewy neurites).

[0235] B8. A compound for use in scheme B7, wherein the use is for brain imaging.

[0236] B9. A compound used for any of schemes B1-B6, for diagnostic purposes.

[0237] B10. A compound for use in scheme B9, for the diagnosis of a disease, condition, or abnormality associated with α-synuclein aggregates (including, but not limited to, Lewy bodies and / or Lewy neurites), wherein said disease, condition, or abnormality is optionally selected from Parkinson's disease (including sporadic, familial with α-synuclein mutations, familial with mutations other than α-synuclein, isolated autonomic dysfunction, or Lewy body dysphagia), Lewy body dementia (LBD), Lewy body dementia (DLB) (including “isolated” Lewy body dementia), Parkinson's disease dementia (PDD), diffuse Lewy body disease (DLBD), sporadic Alzheimer's disease, familial Alzheimer's disease with APP mutations, familial Alzheimer's disease with PS-1, PS-2, or other mutations, familial British dementia, Lewy body variants of Alzheimer's disease, Down syndrome, multiple system atrophy (including Shy-Drager syndrome, striatum, or olive disease). Somatic pontocerebellar atrophy), traumatic brain injury, chronic traumatic encephalopathy, boxer's dementia, tau protein diseases (including Pick's disease, frontotemporal dementia, progressive supranuclear palsy, corticobasal degeneration, Niemann-Pick type C1 disease, frontotemporal dementia associated with Parkinson's syndrome related to chromosome 17), Creutzfeldt-Jakob disease, Huntington's disease, motor neuron disease, amyotrophic lateral sclerosis (including sporadic, familial, or Guam ALS-dementia syndrome), neuroaxial dystrophy, and those with 1 Neurodegenerative diseases caused by iron accumulation in the brain (including Haas-Scheatz syndrome), prions, ataxia-telangiectasia, idiopathic orofacial motor disorders, subacute sclerosing panencephalitis, Gerstmann-Straussler-Scheatz syndrome, inclusion body myositis, Gaucher disease, Krab disease and other lysosomal storage diseases (including Kufor-Rakeb syndrome and Sanfilippo syndrome) and tachyophthalmos (REM) sleep behavior disorders.

[0238] B11. A compound for use in scheme B10, wherein the disease is Parkinson's disease.

[0239] B12. A compound for use in any one of schemes B7-B11, wherein the use is for human use.

[0240] B13. A method for diagnosing a disease, condition, or abnormality, or susceptibility to, α-synuclein aggregates (including but not limited to Lewy bodies and / or Lewy neurites) in a patient, wherein the method comprises:

[0241] a) Administer to the patient a diagnostically effective amount of a compound as defined in any of schemes B1-B6;

[0242] b) Allow the compound to distribute into the tissue of interest; and

[0243] c) Imaging the tissue of interest, wherein increased binding of the compound to the tissue of interest compared to normal control binding levels indicates that the patient has a disease, condition, or abnormality associated with α-synuclein aggregates (including but not limited to Lewy bodies and / or Lewy neurites) or is at risk of developing them.

[0244] B14. A method for collecting data for diagnosing diseases, conditions, or abnormalities in patients associated with α-synuclein aggregates (including but not limited to Lewy bodies and / or Lewy neurites), the method comprising:

[0245] (a) Expose a sample or a specific body part or receptor region suspected of containing α-synuclein aggregates (including but not limited to Lewy bodies and / or Lewy neurites) to a compound as defined in any of schemes B1-B6.

[0246] (b) To bind the compound to α-synuclein aggregates (including but not limited to Lewy bodies and / or Lewy neurites);

[0247] (c) Detecting compounds that bind to α-synuclein aggregates, including but not limited to Lewy bodies and / or Lewy neurites; and

[0248] (d) Optionally establish a correlation between the presence or absence of a compound that binds to α-synuclein aggregates (including but not limited to Lewy bodies and / or Lewy neurites) and the presence or absence of α-synuclein aggregates (including but not limited to Lewy bodies and / or Lewy neurites) in the sample or a specific body part or region.

[0249] B15. Methods for collecting data for diagnosing diseases, conditions, or abnormalities associated with α-synuclein aggregates in patients, including:

[0250] (a) Expose a sample or a specific body part or receptor region suspected of containing α-synuclein aggregates to a compound as defined in any of schemes B1-B6;

[0251] (b) Allowing the compound to bind to the α-synuclein aggregates;

[0252] (c) Detecting compounds that bind to the α-synuclein aggregates; and

[0253] (d) Optionally establish a correlation between the presence or absence of a compound that binds to α-synuclein aggregates and the presence or absence of α-synuclein aggregates in the sample or a specific body part or region.

[0254] B16, a method for determining the amount of α-synuclein aggregates in tissues and / or body fluids, comprising:

[0255] (a) Provide representative tissue and / or body fluid samples for the study;

[0256] (b) Test the presence of α-synuclein aggregates in the sample using a compound as defined in any of schemes B1-B6;

[0257] (c) Determine the amount of compounds that bind to α-synuclein aggregates; and

[0258] (d) Calculate the amount of α-synuclein aggregates in tissues and / or body fluids.

[0259] B17. A method for collecting data to determine susceptibility in patients to diseases, conditions, or abnormalities associated with α-synuclein aggregates, comprising detecting the specific binding of a compound as defined in any of protocols B1-B6 to α-synuclein aggregates in a sample or a specific body site or region, the method comprising the following steps:

[0260] (a) Expose a sample or a specific body part or region suspected of containing α-synuclein aggregates to a compound as defined in schemes B1-B6.

[0261] (b) Allow the compound to bind to α-synuclein aggregates to form a compound / α-synuclein aggregate complex;

[0262] (c) Detection of the formation of the compound / α-synuclein aggregate complex;

[0263] (d) Optionally, establish a correlation between the presence or absence of the compound / α-synuclein aggregate complex and the presence or absence of α-synuclein aggregates in the sample or a specific body part or region; and

[0264] (e) Optionally compare the amount of the compound / α-synuclein aggregate with the normal control value.

[0265] B18. A method for collecting data for monitoring residual disease, condition, or abnormality associated with α-synuclein aggregates that has been treated with medication, wherein the method comprises:

[0266] (a) Expose a sample or a specific body part or region suspected of containing α-synuclein aggregates to a compound as defined in schemes B1-B6.

[0267] (b) Allow the compound to bind to α-synuclein aggregates to form a compound / α-synuclein aggregate complex;

[0268] (c) Detection of the formation of the compound / α-synuclein aggregate complex;

[0269] (d) Optionally, establish a correlation between the presence or absence of the compound / α-synuclein aggregate complex and the presence or absence of α-synuclein aggregates in the sample or a specific body part or region; and

[0270] (e) Optionally compare the amount of the compound / α-synuclein aggregate with the normal control value.

[0271] B19. A method for collecting data to predict the responsiveness of patients with a disease, condition, or abnormality associated with α-synuclein aggregates that are being treated with medication, comprising:

[0272] (a) Expose a sample or a specific body part or region suspected of containing α-synuclein aggregates to a compound as defined in schemes B1-B6.

[0273] (b) Allow the compound to bind to α-synuclein aggregates to form a compound / α-synuclein aggregate complex;

[0274] (c) Detection of the formation of the compound / α-synuclein aggregate complex;

[0275] (d) Optionally, establish a correlation between the presence or absence of the compound / α-synuclein aggregate complex and the presence or absence of α-synuclein aggregates in the sample or a specific body part or region; and

[0276] (e) Optionally compare the amount of the compound / α-synuclein aggregate with the normal control value.

[0277] B20. A diagnostic composition comprising a compound of any one of schemes B1-B6 and a pharmaceutically acceptable excipient, carrier, diluent, or adjuvant.

[0278] B21. Compounds of formula (II-F)

[0279]

[0280] Or its stereoisomers, racemic mixtures, pharmaceutically acceptable salts, hydrates, or solvates.

[0281] in

[0282] R 3 The following are pyrrolidines substituted with leaving groups (LG).

[0283] R 4 It is a 5- or 6-membered heteroaryl group containing one or two N atoms, wherein the heteroaryl group is optionally substituted with a methyl group, and

[0284] * indicates the location of the bond.

[0285] B22. Compounds of formula (II-F) of scheme B21, wherein LG is selected from halogens, C 1-4 Alkyl sulfonates and C 6-10 Aryl sulfonates.

[0286] B23. Compounds of formula (II-F) of schemes B21 or B22, which are

[0287]

[0288] B24. Compounds of formula (II-H)

[0289]

[0290] Or its stereoisomers, racemic mixtures, pharmaceutically acceptable salts, hydrates or solvates, wherein

[0291] R 5 The following are fluorinated pyrrolidines

[0292] R 6 It is a 5- or 6-membered heteroaryl group containing one or two N atoms, wherein the heteroaryl group is optionally substituted with a methyl group and / or the heteroaryl group is optionally substituted with one or more X atoms.

[0293] X is a halogen or H, provided that at least one X is a halogen, and

[0294] * indicates the location of the bond.

[0295] B25. A compound of formula (II-H) of scheme B24, which is

[0296]

[0297] B26. A method for preparing the compound of schemes B2, B3, or B4, comprising mixing the compound of any one of schemes B21-B23 with... 18 The F-fluorinating agent reaction causes LG to be...18 F substitution.

[0298] B27. The method of scheme B26, in which... 18 F-fluorinating agents are selected from K 18 F, H 18 F, Cs 18 F, Na 18 F and [ 18 F]Tetrabutylammonium fluoride.

[0299] B28. A method for preparing the compound of schemes B2, B3, or B5, comprising mixing the compound of any one of schemes B24 or B25 with... 3 H radiolabeled reagent reaction.

[0300] B29. Use of any compound from schemes B1-B6 as an in vitro analytical reference or in vitro screening tool.

[0301] B30. A test kit for detecting and / or diagnosing diseases, conditions or abnormalities associated with α-synuclein aggregates, wherein the test kit contains at least one compound as defined in any one of schemes B1-B6.

[0302] B31. A kit for preparing radiopharmaceutical formulations, wherein the kit comprises a sealed vial containing at least one compound as defined in any one of schemes B21-B25.

[0303] Within schemes A and B, "heterocyclic group" can refer to a carbocyclic group as defined above, wherein at least one carbon atom is partially replaced by a heteroatom selected, for example, from N, O, or S, or a heteroatom-containing group (e.g., N, O, and / or S). The heterocyclic group can be unsaturated or saturated. It encompasses heteroalkyl and heteroaryl groups. The heterocyclic group can also be fused, bridged, or spirolinked, such as 6-membered bicyclic, 7-membered bicyclic, 8-membered bicyclic, 6-membered spirocyclic, 7-membered spirocyclic, or 8-membered spirocyclic. Examples include nitrogen-containing heterocyclic butanes, pyrrolidines, pyrroles, and tetrahydrofurans. Furan, Thiazole pentane, Thiophene, Imidazolidine, Pyrazolidine, Imidazole, Pyrazole, Oxazolidine, Isoxazolidine, Oxazole, Isoxazole, Thiazolidine, Isothiazolidine, Thiazole, Isothiazolidine, Dioxazolidine, Dithiazole, Triazole, Furazan, Oxadiazole, Thiadiazole, Dithiazole, Tetraazole, Piperidine, Oxane, Thiane, Pyridine, Pyran, Thian, Piperazine, Diazine (including pyrazine and pyrimidine), Morpholine, Oxazine, Thiomorpholine Thiazides, dioxanes, dioxins, dithiine, triazine, trioxane, tetraazine, azacycloheptane, acrylonitrile, oxacycloheptane, oxaheptane, thioheptanane, thiaheptanane, 3-azabicyclo[3.1.0]hexane, azaspiro[3.3]heptane, diazaspiro[3.3]heptane, azabicyclo[3.2.1]octane, and diazabicyclo[3.2.1]octane. Preferred examples of heterocyclic groups include azacyclobutane, morpholine, piperazine, pyrrolidine, tetrahydrofuran, piperidine, azaspiro[3.3]heptane, etc. Possible examples of heteroaryl groups include pyridine, pyrazole, etc.

[0304] Regarding these schemes A and B, the following preferred definitions can be applied.

[0305] Preferably, R 2 for

[0306] More preferably, R 2 for

[0307] Even more preferably, R 2 for

[0308] In each of the above implementation schemes, R 2 It can be optionally substituted with methyl.

[0309] F is preferred 19 F or 18 F, more preferred 18 F.

[0310] In one embodiment of schemes A and B, the compound of formula (I) is a detectable labeled compound.

[0311]

[0312] in

[0313] It can detect radioactive isotopes.

[0314] R 1 The following are fluorinated pyrrolidines

[0315] R 2 It is a 5- or 6-membered heteroaryl group containing one or two N atoms, wherein the heteroaryl group is optionally substituted with a methyl group, and

[0316] * indicates the location of the bond.

[0317] Preferably, the detectable marker is a radioactive isotope selected from... 18 F 、 2H and 3 H, optimal choice 18 F and 3 H.

[0318] In one embodiment of schemes A and B, the compound of formula (I) is a detectable labeled compound of formula (IF).

[0319]

[0320] R 1 For the following 18 F-substituted pyrrolidines

[0321] R 2 It is a 5- or 6-membered heteroaryl group containing one or two N atoms, wherein the heteroaryl group is optionally substituted with a methyl group, and

[0322] * indicates the location of the bond.

[0323] In one embodiment of schemes A and B, the compound of formula (I) is a detectable labeled compound of formula (IH).

[0324]

[0325] It is in at least one available location 2 H or 3 H (tritium), preferred 3 H can detect ground markers,

[0326] R 1 The following are fluorinated pyrrolidines

[0327] R 2It is a 5- or 6-membered heteroaryl group containing one or two N atoms, wherein the heteroaryl group is optionally substituted with a methyl group.

[0328] Fluorine is 19 F, and

[0329] * indicates the location of the bond.

[0330] Preferably, the detectable labeled compound of formula (IH) is a compound of formula (I-Ha).

[0331]

[0332] in

[0333] R 1 The following are fluorinated pyrrolidines

[0334] R 2 It is a 5- or 6-membered heteroaryl group containing one or two N atoms, wherein the heteroaryl group is optionally substituted with a methyl group, and / or the heteroaryl group is optionally substituted with at least one T, where T is... 3 H (tritium),

[0335] n is 0-3,

[0336] The condition is that the compound of formula (I-Ha) contains at least one T, where T is... 3 H (tritium),

[0337] Fluorine is 19 F, where * represents the bond position.

[0338] Preferably, the detectable labeled compound of formula (I-Ha) contains one or two Ts.

[0339] Preferably, n is 1.

[0340] In another embodiment, compound R of formula (IH) 2 It is a 6-membered heteroaryl group containing one N atom, wherein the heteroaryl group is substituted by one or more T atoms. Preferably, R2 is... More preferably, R 6 for

[0341] In a preferred embodiment of schemes A and B, the compound of formula (I) is

[0342]

[0343] It may be able to detect labeled compounds, stereoisomers, racemic mixtures, pharmaceutically acceptable salts, hydrates, or solvates, among which...

[0344] R1 The following are fluorinated pyrrolidines

[0345] R 2 It is a 6-membered heteroaryl group containing one or two N atoms, wherein the heteroaryl group is optionally substituted with a methyl group, and

[0346] * indicates the location of the bond.

[0347] Preferably, R 2 It is a 6-membered heteroaryl group containing one nitrogen atom. More preferably, R 2 for

[0348] In the above R 2 In each of the embodiments, the 6-membered heteroaryl group may optionally be substituted with a methyl group.

[0349] definition

[0350] For the purposes of interpreting this specification, the following definitions shall apply unless otherwise stated, and where appropriate, terms used in the singular shall also include the plural, and vice versa.

[0351] "Alkyl" refers to a saturated straight-chain or branched organic moiety consisting of carbon and hydrogen atoms. Alkyl groups typically lack any degree of saturation and are usually linked to the rest of the molecule by single bonds. Suitable examples of alkyl groups have 1 to 6 carbon atoms, preferably 1 to 4 carbon atoms. The term "C1-C4 alkyl" should be interpreted accordingly. Examples of "C1-C4 alkyl" include, but are not limited to, methyl, ethyl, propyl, isopropyl, 1-methylethyl, n-butyl, tert-butyl, and isobutyl.

[0352] "C1-C4 alkoxy" refers to a group having the formula -ORa, where Ra is a C1-C4 alkyl group as generally defined above. Examples of C1-C4 alkoxy groups include, but are not limited to, methoxy, ethoxy, propoxy, isopropoxy, butoxy, and isobutoxy.

[0353] "Halogenated C1-C4 alkyl" or "halogenated C1-C4 alkyl" refers to a C1-C4 alkyl group as defined above, which is substituted by one or more halogen groups as defined below. Examples of "halogenated C1-C4 alkyl" include, but are not limited to, trifluoromethyl, difluoromethyl, fluoromethyl, trichloromethyl, 2,2,2-trifluoroethyl, 1,3-dibromopropyl-2-yl, 3-bromo-2-fluoropropyl, and 1,4,4-trifluorobutyl-2-yl.

[0354] "C3-C6 cycloalkyl" refers to a stable monocyclic saturated hydrocarbon group consisting of 3 to 6 carbon atoms, composed only of carbon and hydrogen atoms. Examples of C3-C6 cycloalkyl include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, and cyclohexyl.

[0355] A "heterocyclic group" refers to a stable 4- to 6-membered non-aromatic monocyclic group containing one or two heteroatoms, for example, selected from N, O, or S. The heterocyclic group can be unsaturated or saturated. The heterocyclic group can be bonded by carbon atoms or heteroatoms. Examples include, but are not limited to, azirrocyclobutane, oxoheterocyclobutane, pyrrolinyl, pyrrolylyl, tetrahydrofuranyl, tetrahydrothiophenyl, piperidinyl, piperazinyl, tetrahydropyranyl, morpholinyl, or perhydroazirrocyclobutane. Preferred examples of heterocyclic groups include, but are not limited to, azirrocyclobutane, morpholinyl, piperazinyl, pyrrolylyl, or piperidinyl.

[0356] "Aryl" refers to a carbocyclic aromatic organic moiety composed of carbon and hydrogen atoms (e.g., containing one or two rings), preferably having 5 to 12 carbon atoms, more preferably 6 to 12 carbon atoms, more preferably 6 to 10 carbon atoms, even more preferably 5 to 10 carbon atoms, and even more preferably 5 or 6 carbon atoms. Examples include, but are not limited to, phenyl, biphenyl, and naphthyl.

[0357] "Heteroaryl" refers to an aryl group as defined above, wherein at least one carbon atom is partially substituted by a heteroatom selected, for example, from N, O, or S, or a heteroatom-containing group (e.g., N, O, and / or S). Typically, heteroaryl groups are 5- to 8-membered ring systems, preferably 5- to 6-membered ring systems, wherein at least one carbon atom is substituted by a heteroatom selected, for example, from N, O, or S. Examples of possible heteroaryl groups include, but are not limited to, furanyl, pyrroleyl, thiopheneyl, pyrazolyl, imidazolyl, thiazolyl, isothiazolyl, oxazolyl, isoxazolyl, triazolyl, tetrazolyl, pyrazinyl, pyridazinyl, pyrimidinyl, or pyridinyl. Preferred examples include pyridine, pyrazole, etc., more preferably pyridine.

[0358] "Hal" or "halogenated" refers to F, Cl, Br, and I. In diagnostic and pharmaceutical applications, F (e.g., Br) is particularly preferred. 19 F and 18 F).

[0359] As used herein, the term “leaving group” (LG) is any leaving group and means an atom or group that can be replaced by another atom or group. For example, in Synthesis (1982), pp. 85-125, Table 2; Carey and Sundberg, Organische Synthese, (1995), pp. 279-281, Table 5.8; or Netscher, Recent Res. Dev. Org. Chem., 2003, 7, 71-83, Schemes 1, 2, 10 and 15, etc.). (Coenen, Fluorine-18 Labeling Methods: Features and Possibilities of Basic Reactions, (2006): Schubiger PA, Friebe M., Lehmann L., (eds), PET-Chemistry-The Driving Force in Molecular Imaging. Springer, Berlin Heidelberg, pp. 15-50, explicitly: Scheme 4 on page 25, Scheme 5 on page 28, Table 4 on page 30, page 33) Figure 7 Examples are given in ( ). Preferably, the "leaving group" (LG) is selected from halogens, C 1-4 Alkyl sulfonates and C 6-10 Aryl sulfonates, wherein C 6-10 The aryl group can be optionally replaced by -CH3 or -NO2.

[0360] Unless otherwise stated, the term "compound of the present invention" refers to a compound of formula (I) or its subforms (e.g., (IIa), (IIb), (IIIa), (IIIb), (IIIc), (III-F), (III-H)) or a compound of which it is detectably labeled, stereoisomers (including mixtures of diastereomers and individual diastereomers, mixtures of enantiomers and individual enantiomers, mixtures of conformational isomers and individual conformational isomers), racemic mixtures, pharmaceutically acceptable salts, hydrates, or solvates. It should be understood that each reference to a compound of formula (I) as defined herein also encompasses its subforms (e.g., (IIa), (IIb), (IIIa), (IIIb), (IIIc), (III-F), (III-H)).

[0361] The compounds of the present invention and their precursors having one or more optically active carbons can exist as racemates and mixtures of racemates, stereoisomers (including mixtures of diastereomers and individual diastereomers, mixtures of enantiomers and individual enantiomers, mixtures of conformational isomers and individual conformational isomers), tautomers, transisomers, and rotational isomers. All isomer forms are included in this invention. Compounds containing alkene double bonds described in this specification include E and Z geometrical isomers.

[0362] The invention also includes all salt forms, polymorphs, hydrates, and solvates (e.g., ethanolates).

[0363] A "pharmaceutically acceptable salt" is defined as a derivative of the disclosed compound, wherein the parent compound is modified by preparing its acid salt or base salt. Examples of pharmaceutically acceptable salts include, but are not limited to, inorganic or organic acid salts of basic groups such as amines; alkali metal salts or organic salts of acidic groups such as carboxylic acids. Pharmaceutically acceptable salts include, for example, conventional non-toxic salts or quaternary ammonium salts of parent compounds formed from non-toxic inorganic or organic acids. For example, such conventional non-toxic salts include those derived from inorganic acids, such as, but not limited to, hydrochloric acid, hydrobromic acid, sulfuric acid, aminosulfonic acid, phosphoric acid, nitric acid, etc.; and salts prepared from organic acids, such as, but not limited to, acetic acid, propionic acid, succinic acid, glycolic acid, stearic acid, lactic acid, malic acid, tartaric acid, citric acid, ascorbic acid, dihydroxynaphthyl acid, maleic acid, hydroxymaleic acid, phenylacetic acid, glutamic acid, benzoic acid, salicylic acid, p-aminobenzenesulfonic acid, 2-acetoxybenzoic acid, fumaric acid, toluenesulfonic acid, methanesulfonic acid, ethanedisulfonic acid, oxalic acid, hydroxyethanesulfonic acid, etc. Pharmaceutically acceptable salts of the compounds of the present invention and their precursors can be synthesized by conventional chemical methods from parent compounds containing a basic or acidic moiety. Typically, such salts can be prepared by reacting the free acidic or basic form of these compounds with a stoichiometric amount of a suitable base or acid in water, in an organic solvent, or in a mixture of both. Organic solvents include, but are not limited to, non-aqueous media such as ethers, ethyl acetate, ethanol, isopropanol, or acetonitrile. A list of suitable salts can be found in Remington's Pharmaceutical Sciences, 18th edition, Mack Publishing Company, Easton, PA, 1990, p. 1445, the contents of which are incorporated herein by reference.

[0364] "Pharmaceutical acceptable" is defined as those compounds, materials, compositions, and / or dosage forms that, within reasonable medical judgment, are suitable for use in contact with human and animal tissues without excessive toxicity, irritation, allergic reactions, or other problems or complications, in proportion to a reasonable benefit / risk ratio.

[0365] The compounds of the present invention may also be provided in the form of prodrugs, i.e., compounds that are metabolized in vivo into active metabolites.

[0366] The patients or individuals in this invention are typically animals, particularly mammals, and even more particularly humans.

[0367] Alpha-synuclein aggregates are β-sheet-rich polymeric assemblages of α-synuclein monomers that can form soluble oligomers or soluble / insoluble primary or mature fibrils. They aggregate into intracellular deposits and are detected as a series of Lewy lesions in Parkinson's disease and other synucleinopathies. Alpha-synuclein aggregates constituting Lewy pathology can be detected with the following morphologies: Lewy bodies, Lewy neurites, precocious Lewy bodies or pallid bodies, and perinuclear deposits with diffuse, granular, punctate, or pleomorphic patterns. Furthermore, α-synuclein aggregates are a major component of intracellular fibrillary inclusions detected in oligodendrocytes (also known as glial cytoplasmic inclusions) and neuronal somatic cells, axons, and nuclei (called neuronal cytoplasmic inclusions), and are a histological marker of multiple system atrophy. In Lewy lesions, α-synuclein aggregates typically show a significant increase in post-translational modifications such as phosphorylation, ubiquitination, nitration, and truncation.

[0368] Lewy bodies are abnormal aggregates of proteins that develop within nerve cells in Parkinson's disease (PD), Lewy body dementia, and other synucleinopathies. Lewy bodies appear as spherical clumps that replace other cellular components. Morphologically, Lewy bodies can be classified as brainstem or cortical types. Typical brainstem Lewy bodies are eosinophilic cytoplasmic inclusions consisting of a dense core surrounded by a halo of radiating fibrils 5-10 nm wide, with α-synuclein as their primary structural component; cortical Lewy bodies differ in that they lack the halo. The presence of Lewy bodies is a hallmark of Parkinson's disease.

[0369] Lewy neurites are abnormal neuronal processes in diseased neurons, containing granular material, abnormal α-synuclein filaments similar to those found in Lewy bodies, punctate, varicose structures, and axonal spheroids. Like Lewy bodies, Lewy neurites are characteristic of α-synucleinopathies such as Lewy body dementia, Parkinson's disease, and multiple system atrophy.

[0370] The terms “disease,” “symptom,” or “abnormality” are used interchangeably in this article.

[0371] Compounds of Formula (I) or their detectable labeled compounds, stereoisomers, racemic mixtures, pharmaceutically acceptable salts, hydrates, or solvates can bind to α-synuclein aggregates. The type of bond between compounds of Formula (I) or their detectable labeled compounds, stereoisomers, racemic mixtures, pharmaceutically acceptable salts, hydrates, or solvates has not been elucidated, and this invention covers any type of bond. The terms "compounds binding to α-synuclein aggregates," "compound / (α-synuclein aggregate (including but not limited to Lewy bodies and / or Lewy neurites)) complex," "compound / α-synuclein aggregate complex," "compound / protein aggregate complex," etc., are used interchangeably herein and are not considered to be limited to any particular type of bond.

[0372] Unless otherwise stated, the preferred definitions given in the "Definitions" section apply to all embodiments described below. Various embodiments of the invention are described herein, and it should be understood that features specified in each embodiment can be combined with other specified features to provide further embodiments of the invention. Brief description of the attached diagram

[0374] Figure 1 [3H]-Example-1 / Example-1[ 3 [H-1] targets binding on tissues from various α-synucleinopathies. Accumulation of silver granules on Lewy bodies and Lewy neurites is shown in the lower figure. Immunofluorescence staining with α-syn-pS129 antibody on the same sections is shown in the upper figure to co-label α-syn aggregates. PD, Parkinson's disease; PDD, Parkinson's disease with dementia; MSA, multiple system atrophy; DLB, Lewy body dementia; LBV, Lewy body variant of Alzheimer's disease. Scale bar, 20 μm.

[0375] Figure 2 Evaluation of Example 1 by Autoradiography 3 [H-1] binding affinity to human PDD brain tissue. A) Autoradiography image, B) Immunofluorescence staining with α-syn-pS129 antibody, C) Example-1 [ 3 Specific binding of H-1] (RU: relative unit). Scale bar, 2 mm. '-', total binding; '+', self-closing, nonspecific binding.

[0376] Figure 3 Evaluation of Example 1 by Autoradiography 3 [H-1] Binding affinity to human brain tissue from familial PD cases (G51D missense mutation). A) Autoradiography image, B) Immunofluorescence staining with α-Syn-pS129 antibody, C) Example-1 [3 Specific binding of H-1] (RU: relative unit). Scale bar, 5 mm. '-', total binding; '+', self-closing, nonspecific binding.

[0377] Figure 4 Example 1: Evaluation by Autoradiography 3 [H-1] binding specificity and head-to-head comparison with the reference α-syn binder ([3H]-α-syn-ref). A) Autoradiographic image, B) Immunofluorescence staining with α-SYN-pS129 antibody. Scale bar, 2 mm. PDD, Parkinson's disease with dementia; PD_SNCA, G51D missense mutation in α-synuclein [SNCA] gene; NDC, non-dementia control. '-', total binding; '+', self-blocking, non-specific (NS) binding.

[0378] Figure 5 The saturation binding of [3H]-Example 1 to α-syn aggregates from PD brain was tested by microradiobinding assay and compared head-to-head with [3H]-α-syn-Ref. The figure shows the specific binding (RU: relative unit).

[0379] Figure 6 Example 1 3 H-1] competitively binds to α-syn-Ref on α-syn aggregates derived from the brain in idiopathic PD. Example-1 [ 3 The percentage of competition for H-1] is plotted relative to the concentration of the non-radiolabeled α-syn-Ref (left) or the compound of Example 1 (right). The average value of the two technical replicates is shown.

[0380] Figure 7 Evaluation of the Ki value of the compound of Example 1 when the reference compound Aβ ([3H]-Abeta-Ref) was replaced with the non-radioactively labeled compound of Example 1 in the homogenate of AD brain-derived AD. 3 A graph was plotted showing the percentage increase in the competition for H]-Aβ-Ref binding relative to the concentration of the non-radiolabeled compound from Example 1. The average value of the two technical replicates is shown.

[0381] Figure 8 Example 1 3 [H-1] Evaluation of target binding in AD tissues containing pathological Tau aggregates. A) Immunofluorescence staining of the same tissues labeled with Tau aggregates using MCI antibody; B) Comparison with a reference Tau ligand ([3H]-Tau-ref) using Example-1 [ 3 [H-1] No silver particles accumulated on the Tau entanglement.

[0382] Figure 9 Example 1 3 [H-1] Evaluation of target binding in frontotemporal degeneration (FTLD) TDP type C tissue containing pathological TDP-43 aggregates. Immunofluorescence staining with phosphate-TDP-43 antibody was performed on the same tissue-labeled TDP-43 aggregates (top figure). In use in Example-1 [ 3 No silver particles accumulated on the TDP-43 aggregates of H-1] (bottom figure). Scale bar, 20 μm.

[0383] Figure 10 Example 1-[ 18 F-1]] IV NHP PK was performed throughout the monkey brain.

[0384] Figure 11 The binding specificity of Example 1 [3H-1] to different α-synucleinopathies and non-dementia controls (NDCs) was evaluated by autoradiography. A) Autoradiographic images; B) Immunofluorescence staining of diseased donors with α-syn-pS129 antibody. Scale bar, 5 mm. PDD, Parkinson's disease with dementia; MSA, multiple system atrophy; LBV, Lewy body variant of Alzheimer's disease; NDC, non-dementia controls. 'Total', total binding; 'NSB', non-specific binding.

[0385] Figure 12 :[ 3 H]-Example-4 / Example-4[ 3 H-4] target binding on PD tissue. Accumulation of silver particles on Lewy bodies and Lewy neurites, as shown in the lower figure. Immunofluorescence staining with α-syn-pS129 antibody on the same sections, as shown in the upper figure, to co-label α-syn aggregates. Scale bar, 20 μm.

[0386] Figure 13 Example 4 of autoradiography 3 [H-4] Assessment of binding specificity in different α-synucleinemia and non-dementia control cases. A) Autoradiographic images; B) Immunofluorescence staining of diseased donors with α-syn-pS129 antibody. Scale bar, 2 mm. SNCA, G51D missense mutation in α-synuclein [SNCA] gene; PD, Parkinson's disease; MSA, multiple system atrophy; NDC, non-dementia control. 'Total', total binding; 'NSB', non-specific binding.

[0387] Figure 14 :By microradioactive binding to α-syn aggregates derived from PD brain [ 3 H] - Saturated binding of Example 4. This figure shows the specific binding (per mm).2 (Count per minute). The mean (mean ± SD) of the four independent experiments is shown.

[0388] Figure 15 The reference Aβ compound was replaced with the non-radioactively labeled compound of Example 4 on an AD brain-derived homogenate. 3 Evaluation of the Ki value of the compound in Example 4 (H]-Abeta-Ref). 3 The percentage of competing H]-Aβ-Ref bindings was plotted relative to the concentration of the non-radiolabeled compound in Example 4. The average (mean ± SD) of two independent experiments with two technical replicates is shown.

[0389] Figure 16 ： Example 4 was performed using microautoradiography. 3 Evaluation of target binding of [H-4] in AD tissues containing pathological Tau aggregates. Compared with reference Tau ligand ([ 3 Compared to H]-Tau-ref), using Example-4[ 3 No silver particle accumulation was observed on the Tau tangles of H-1]. Invention Details

[0391] The compounds and precursors of the present invention are described below. It should be understood that all possible combinations of the following definitions are also covered.

[0392] This invention relates to compounds of formula (I),

[0393]

[0394] Or its detectable labeled compounds, stereoisomers, racemic mixtures, pharmaceutically acceptable salts, hydrates, or solvates, wherein

[0395] It is an aryl or heteroaryl group, oriented and selected from the following:

[0396]

[0397] R 0 It is H or C1-C4 alkyl;

[0398] R 1 -CN; or halogen; or C1-C4 alkyl; or C1-C4 alkoxy; or -N(C1-C4 alkyl)2; or -NH(C1-C4 alkyl); or H, or

[0399] R 1 It is a -NH-C3-C6 cycloalkyl, C3-C6 cycloalkyl or heterocyclic group, each of which is optionally substituted with at least one halogen;

[0400] R 2 It is an aryl, 5-membered, or 6-membered heteroaryl group, wherein R 2 Selected from the following:

[0401]

[0402] in

[0403] R 2a R 2a’ Independently selected from H or F;

[0404] R 2b Independently selected from F, -OH, C1-C4 alkyl, halo-C1-C4 alkyl, -NH2, -CN or C1-C4 alkoxy;

[0405] R 2c R 2c’ It is independently selected from H, F, OH, OCH3 or CH3;

[0406] R 2d Selected from H, F, or -OH;

[0407] R 2e Selected from H, OH, CH3 or F;

[0408] Z can be N, NH, N (C1-C4 alkyl), N (halogenated C1-C4 alkyl), O, or S independently;

[0409] Z 1 It can be N, NH, O, or S independently;

[0410] p is 0, 1, or 2;

[0411] m is 0 or 1;

[0412] When valence is allowed It is a combination of single and double bonds; and

[0413] * indicates the location of the bond.

[0414] In another embodiment, the present invention provides a compound of formula (I) having formula (IIa) or (IIb).

[0415]

[0416] Or its detectable labeled compounds, stereoisomers, racemic mixtures, pharmaceutically acceptable salts, hydrates, or solvates.

[0417] In another embodiment, the present invention provides compounds of formula (I) having formula (IIIa), (IIIb) or (IIIc),

[0418]

[0419] Or its detectable labeled compounds, stereoisomers, racemic mixtures, pharmaceutically acceptable salts, hydrates, or solvates.

[0420] R 0 It is H or C1-C4 alkyl. Preferably, R 0 H or CH3, more preferably, R 0 For H.

[0421] In one implementation, R 1 It is H, -CN, halogen, C1-C4 alkyl, C1-C4 alkoxy, N(C1-C4 alkyl)2, or -NH(C1-C4 alkyl). Preferably, R 1 It is -CN, halogen, C1-C4 alkyl, C1-C4 alkoxy, N(C1-C4 alkyl)2, or -NH(C1-C4 alkyl). More preferably, R 1 It is -CN, F, C1-C3 alkyl, C1-C3 alkoxy, or -N(C1-C3 alkyl)2. Even more preferably, R 1 It is -CN, -CH(CH3)2, -OCH3, -OCH(CH3)2, -N(CH3)2 or -NH-CH(CH3)2.

[0422] In one implementation, R 1 It is an -NH-C3-C6 cycloalkyl, C3-C6 cycloalkyl, or heterocyclic group, each optionally substituted with at least one halogen. Preferably, R 1 Selected from the following:

[0423]

[0424] Where R 1’ It is a halogen on its own; and s = 0, 1, 2 or 3.

[0425] More preferably, R 1 Selected from the following:

[0426]

[0427] Even more preferably, R 1 Selected from

[0428]

[0429] In a preferred embodiment, F is preferably... 19 F or 18 F, more preferred 18 F.

[0430] In one implementation, R 2 Selected from the following:

[0431]

[0432] in

[0433] R 2a R2 a’ Independently selected from H or F;

[0434] R 2b Independently selected from F, -OH, C1-C4 alkyl, halo-C1-C4 alkyl, -NH2, -CN or C1-C4 alkoxy;

[0435] R 2c R 2c’ It is independently selected from H, F, OH, OCH3 or CH3;

[0436] R 2d Selected from H, F, or -OH;

[0437] R 2e Selected from H, OH, CH3 or F;

[0438] Z can be N, NH, N (C1-C4 alkyl), N (halogenated C1-C4 alkyl), O, or S independently;

[0439] Z 1 It can be N, NH, O, or S independently;

[0440] p is 0, 1, or 2;

[0441] m is 0 or 1;

[0442] When valence is allowed It is a combination of single and double bonds; and

[0443] * indicates the location of the bond.

[0444] Preferably, R 2 Selected from the following:

[0445]

[0446] in

[0447] R 2a R 2a’ Independently selected from H or F;

[0448] R 2b Independently selected from F, -OH, C1-C4 alkyl, halo-C1-C4 alkyl, -NH2, -CN or C1-C4 alkoxy;

[0449] R 2c R 2c’ It is independently selected from H, F, OH, OCH3 or CH3;

[0450] R 2d Selected from H, F, or -OH;

[0451] R 2e Selected from H, OH, CH3 or F;

[0452] R z Selected from H, C1-C4 alkyl, and halo-C1-v4 alkyl;

[0453] p is 0, 1, or 2; and

[0454] * indicates the location of the bond.

[0455] Preferably, R 2 Selected from the following:

[0456]

[0457] in

[0458] R 2a R 2a’ Independently selected from H or F;

[0459] R 2b Independently selected from F, -OH, C1-C4 alkyl, halo-C1-C4 alkyl, -NH2, -CN or C1-C4 alkoxy;

[0460] R 2c R 2c’ It is independently selected from H, F, OH, OCH3 or CH3;

[0461] R 2d Selected from H, F, or -OH;

[0462] R 2e Selected from H, OH, CH3 or F;

[0463] R z Selected from H, C1-C4 alkyl, and halo-C1-C4 alkyl;

[0464] p is 0, 1, or 2; and

[0465] * indicates the location of the bond.

[0466] More preferably, R 2 Selected from the following:

[0467]

[0468] Where R 2aR 2a’ Independently selected from H or F;

[0469] R 2b Independently selected from F, -OH, C1-C4 alkyl, halo-C1-C4 alkyl, -NH2, -CN or C1-C4 alkoxy;

[0470] R 2c R 2c’ It is independently selected from H, F, OH, OCH3 or CH3;

[0471] R 2e Selected from H, OH, CH3 or F;

[0472] R z Selected from H, C1-C4 alkyl, and halo-C1-C4 alkyl;

[0473] p is 0, 1, or 2; and

[0474] * indicates the location of the bond.

[0475] Even more preferably, R2 is selected from:

[0476] The asterisk (*) indicates the location of the bond.

[0477] In another embodiment, the present invention provides a compound of either formula (IIa) or (IIb).

[0478]

[0479] Or its detectable labeled compounds, stereoisomers, racemic mixtures, pharmaceutically acceptable salts, hydrates, or solvates, wherein R 0 Methyl or H; R 1 It is CH3 or H; preferably, R 1 It is CH3; and R 2 Contains at least one fluorine fluorine, preferably from the following:

[0480]

[0481] in

[0482] R 2a R 2a’ Independently selected from H or F;

[0483] R 2b Independently selected from F, -OH, C1-C4 alkyl, halo-C1-C4 alkyl, -NH2, -CN or C1-C4 alkoxy;

[0484] R 2c R 2c’It is independently selected from H, F, OH, OCH3 or CH3;

[0485] R 2d Selected from H, F, or -OH;

[0486] R 2e Selected from H, OH, CH3 or F;

[0487] R z Selected from H, C1-C4 alkyl, and halo-C1-C4 alkyl;

[0488] p is 0, 1, or 2; and

[0489] * indicates the location of the bond.

[0490] Most preferably, R 2 Selected from

[0491]

[0492] Where R 2a R 2a’ R 2b R 2e R 2c R 2c’ R z And p as defined above; and where R 2a R 2a’ R 2b R 2c R 2c 'and R 2e At least one of them is F. F is preferably... 19 F or 18 F, more preferred 18 F.

[0493] In another embodiment, the present invention provides a compound of any one of sub-forms (IIIa), (IIIb), or (IIIc).

[0494]

[0495] Or its detectable labeled compounds, stereoisomers, racemic mixtures, pharmaceutically acceptable salts, hydrates, or solvates.

[0496] Where R 0 It is methyl or H, preferably R 0 For H;

[0497] R 1 Selected from -CN, halogen, C1-C4 alkyl; or C1-C4 alkoxy, -N(C1-C4 alkyl)2, -NH(C1-C4 alkyl), H; or

[0498] R 1 It is a -NH-C3-C6 cycloalkyl, C3-C6 cycloalkyl or heterocyclic group, each of which is optionally substituted with at least one halogen;

[0499] Preferably, R 1 Selected from the following:

[0500]

[0501] F is preferred 19 F or 18 F, more preferred 18 F; and

[0502] R 2 The following are preferred:

[0503]

[0504] in

[0505] R 2a R 2a’ Independently selected from H or F;

[0506] R 2b Independently selected from F, -OH, C1-C4 alkyl, halo-C1-C4 alkyl, -NH2, -CN or C1-C4 alkoxy;

[0507] R 2c R 2c’ It is independently selected from H, F, OH, OCH3 or CH3;

[0508] R 2d Selected from H, F, or -OH;

[0509] R 2e Selected from H, OH, CH3 or F;

[0510] R z Selected from H, C1-C4 alkyl, and halo-C1-C4 alkyl;

[0511] p is 0, 1, or 2; and

[0512] * indicates the location of the bond.

[0513] In another embodiment, the present invention relates to compounds of formula (IIIa):

[0514]

[0515] Or its detectable labeled compounds, stereoisomers, racemic mixtures, pharmaceutically acceptable salts, hydrates, or solvates, wherein

[0516] R0 It is methyl or H, preferably R 0 For H;

[0517] R 1 It is a -NH-C3-C6 cycloalkyl, C3-C6 cycloalkyl, or heterocyclic group, each optionally substituted with at least one halogen, preferably, R 1 Selected from the following:

[0518]

[0519] R 1 Preferably, the following are substituted with fluorine:

[0520] More preferably, R 1 for Preferably, R 1 for

[0521] R 2 The following are preferred:

[0522]

[0523] in

[0524] R 2a R 2a’ Independently selected from H or F;

[0525] R 2b Independently selected from F, -OH, C1-C4 alkyl, halo-C1-C4 alkyl, -NH2, -CN or C1-C4 alkoxy;

[0526] R 2c R 2c’ It is independently selected from H, F, OH, OCH3 or CH3;

[0527] R 2d Selected from H, F, or -OH;

[0528] R 2e Selected from H, OH, CH3 or F;

[0529] R z Selected from H, C1-C4 alkyl, and halo-C1-C4 alkyl;

[0530] p is 0, 1, or 2; and

[0531] * indicates the location of the bond.

[0532] Preferably, R 2 Selected from the following:

[0533]

[0534] Where R 2a R 2a’ R 2b R 2c R 2c’ R 2d R 2e R z p is as defined above.

[0535] More preferably, R 2 Selected from the following:

[0536]

[0537] Where R 2a R 2a’ R 2b R 2c R 2c’ R 2d R 2e R z p is as defined above.

[0538] Preferably, R 2 for

[0539] More preferably, R 2 for

[0540] Even more preferably, R 2 for

[0541] In each of the above implementation schemes, R 2 It may optionally be substituted with one or more substituents as disclosed above. F is preferred. 19 F or 18 F, more preferred 18 F.

[0542] In another embodiment, the present invention relates to compounds of formula (IIIb):

[0543]

[0544] Or its detectable labeled compounds, stereoisomers, racemic mixtures, pharmaceutically acceptable salts, hydrates, or solvates, wherein

[0545] R 0 It is methyl or H, preferably R 0 For H;

[0546] R 1It is a -NH-C3-C6 cycloalkyl, C3-C6 cycloalkyl, or heterocyclic group, each optionally substituted with at least one halogen, preferably, R 1 Selected from the following:

[0547]

[0548] R 1 Preferably, the following are substituted with fluorine:

[0549] More preferably, R 1 for Preferably, R 1 for

[0550] R 2 The following are preferred:

[0551]

[0552] in

[0553] R 2a R 2a’ Independently selected from H or F;

[0554] R 2b Independently selected from F, -OH, C1-C4 alkyl, halo-C1-C4 alkyl, -NH2, -CN or C1-C4 alkoxy;

[0555] R 2c R 2c’ It is independently selected from H, F, OH, OCH3 or CH3;

[0556] R 2d Selected from H, F, or -OH;

[0557] R 2e Selected from H, OH, CH3 or F;

[0558] R z Selected from H, C1-C4 alkyl, and halo-C1-C4 alkyl;

[0559] p is 0, 1, or 2; and

[0560] * indicates the location of the bond.

[0561] Preferably, R 2 Selected from the following:

[0562]

[0563] Where R 2a R 2a’ R 2b R2c R 2c’ R 2d R 2e R z p is as defined above.

[0564] More preferably, R 2 Selected from the following:

[0565]

[0566] Where R 2a R 2a’ R 2b R 2c R 2c’ R 2d R 2e R z p is as defined above.

[0567] Preferably, R 2 for

[0568] More preferably, R 2 for

[0569] Even more preferably, R 2 for

[0570] In each of the above implementation schemes, R 2 It may optionally be substituted with one or more substituents as disclosed above. F is preferred. 19 F or 18 F, more preferred 18 F.

[0571] In another embodiment, the present invention relates to compounds of formula (IIIc):

[0572] Or its detectable labeled compounds, stereoisomers, racemic mixtures, pharmaceutically acceptable salts, hydrates, or solvates, wherein

[0573] R 0 It is methyl or H, preferably R 0 For H;

[0574] R 1 It is a -NH-C3-C6 cycloalkyl, C3-C6 cycloalkyl, or heterocyclic group, each optionally substituted with at least one halogen, preferably, R 1 Selected from the following:

[0575]

[0576] R 1 Preferably, the following are substituted with fluorine:

[0577] More preferably, R 1 for Preferably, R 1 for

[0578] R 2 The following are preferred:

[0579]

[0580] in

[0581] R 2a R 2a’ Independently selected from H or F;

[0582] R 2b Independently selected from F, -OH, C1-C4 alkyl, halo-C1-C4 alkyl, -NH2, -CN or C1-C4 alkoxy;

[0583] R 2c R 2c’ It is independently selected from H, F, OH, OCH3 or CH3;

[0584] R 2d Selected from H, F, or -OH;

[0585] R 2e Selected from H, OH, CH3 or F;

[0586] R z Selected from H, C1-C4 alkyl, and halo-C1-C4 alkyl;

[0587] p is 0, 1, or 2; and

[0588] * indicates the location of the bond.

[0589] Preferably, R 2 Selected from the following:

[0590]

[0591] Where R 2a R 2a’ R 2b R 2c R 2c’ R 2d R 2e R z p is as defined above.

[0592] More preferably, R 2Selected from the following:

[0593]

[0594] Where R 2a R 2a’ R 2b R 2c R 2c’ R 2d R 2e R z p is as defined above.

[0595] Preferably, R 2 for

[0596] More preferably, R 2 for

[0597] Even more preferably, R 2 for

[0598] In each of the above implementation schemes, R 2 It may optionally be substituted with one or more substituents as disclosed above. F is preferred. 19 F or 18 F, more preferred 18 F.

[0599] In another embodiment, the present invention provides compounds of formula (I) or its subforms (e.g., (IIa), (IIb), (IIIa), (IIIb), (IIIc), (III-F), (III-H)) or detectably labeled compounds thereof, stereoisomers, racemic mixtures, pharmaceutically acceptable salts, hydrates or solvates, wherein preferred compounds are:

[0600]

[0601]

[0602]

[0603]

[0604]

[0605]

[0606]

[0607]

[0608]

[0609] More preferably, the stereoisomer of the preferred compound is

[0610]

[0611] In one embodiment, the present invention provides a compound of formula (I) or its sub-forms (e.g., (IIa), (IIb), (IIIa), (IIIb), (IIIc), (III-F), (III-H)) or its stereoisomers, racemic mixtures, pharmaceutically acceptable salts, hydrates or solvates, wherein the compound of formula (I) is a detectably labeled compound.

[0612] One embodiment of the present invention provides a compound of formula (I) or its sub-forms (e.g., (IIa), (IIb), (IIIa), (IIIb), (IIIc), (III-F), (III-H)) or its stereoisomers, racemic mixtures, pharmaceutically acceptable salts, hydrates or solvates, wherein the compound is a detectably labeled compound, wherein the detectable label is a radioisotope, and wherein the compound of formula (I) contains at least one radioisotope.

[0613] Preferably, the detectable marker is a radioactive isotope selected from... 18 F, 2 H and 3 H, optimal choice 18 F or 3 H.

[0614] In one embodiment, the present invention provides a compound of formula (I), preferably a compound of formula (IIIa), or a detectable labeled compound thereof, stereoisomer, racemic mixture, pharmaceutically acceptable salt, hydrate, or solvate thereof, wherein the compound is a detectable labeled compound of formula (III-F).

[0615]

[0616] Or its stereoisomers, racemic mixtures, pharmaceutically acceptable salts, hydrates or solvates, wherein

[0617] R 1 The following were 18 F replaces:

[0618] R 2 It is an aryl, 5-membered, or 6-membered heteroaryl group, wherein R 2 Selected from

[0619]

[0620] in

[0621] R 2a R 2a’ Independently selected from H or F;

[0622] R 2b Independently selected from F, -OH, C1-C4 alkyl, halo-C1-C4 alkyl, -NH2, -CN or C1-C4 alkoxy;

[0623] R 2c R 2c’ It is independently selected from H, F, OH, OCH3 or CH3;

[0624] R 2d Selected from H, F, or -OH;

[0625] R 2e Selected from H, OH, CH3 or F;

[0626] Z can be N, NH, N (C1-C4 alkyl), N (halogenated C1-C4 alkyl), O, or S independently;

[0627] Z 1 It can be N, NH, O, or S independently;

[0628] p is 0, 1, or 2;

[0629] m is 0 or 1;

[0630] When the valence is allowed It is a combination of single and double bonds; and

[0631] * indicates the location of the bond.

[0632] Preferably, R 2 Selected from

[0633]

[0634] Where R 2a R 2a’ R 2b R 2c R 2c’ R 2d R 2e p is as defined above, and R z Selected from H, C1-C4 alkyl and halogenated C1-C4 alkyl.

[0635] More preferably, R 2 Selected from the following:

[0636]

[0637] Where R 2a R 2a’ R 2b R 2c R 2c’ R 2d R 2e R z p is as defined above.

[0638] More preferably, R 2 Selected from the following:

[0639]

[0640] Where R 2a R 2a’ R 2b R 2c R 2c’ R 2e R z p is as defined above.

[0641] Preferably, the detectable labeled compound of formula (III-F) or its stereoisomers, racemic mixtures, pharmaceutically acceptable salts, hydrates, or solvates contain at least one 18 F. Preferably, R 2 (e.g. R) 2a R 2a’ R 2b R 2c R 2c’ R z and R 2e The substituents of ) can optionally be 18 F. More preferably, the detectable labeled compound of formula (III-F) or its stereoisomers, racemic mixtures, pharmaceutically acceptable salts, hydrates or solvates contain one or two of these compounds. 18 F. Even more preferably, a 18 F.

[0642] Preferred compounds are selected from:

[0643]

[0644] Or its pharmaceutically acceptable salts, hydrates or solvates.

[0645] The most preferred compound is

[0646]

[0647] Or its pharmaceutically acceptable salts, hydrates or solvates.

[0648] In one embodiment, the present invention provides a compound of formula (I), preferably a compound of formula (IIIa) or a detectable labeled compound thereof, a stereoisomer, a racemic mixture, a pharmaceutically acceptable salt, a hydrate, or a solvate, wherein the compound is a detectable labeled compound of formula (III-H).

[0649]

[0650] Or its stereoisomers, racemic mixtures, pharmaceutically acceptable salts, hydrates, or solvates.

[0651] It is in at least one available location 2 H (deuterium "D") or 3 H (tritium "T"), preferred 3 H can detect ground markers,

[0652] in

[0653] R 1 -CN; or halogen; or C1-C4 alkyl; or C1-C4 alkoxy; or -N(C1-C4 alkyl)2; or -NH(C1-C4 alkyl); or H; or

[0654] R 1 It is a -NH-C3-C6 cycloalkyl, C3-C6 cycloalkyl, or heterocyclic group, each optionally substituted with at least one halogen; R 1 Preferred from

[0655]

[0656] R 2 It is an aryl, 5-membered, or 6-membered heteroaryl group, wherein R 2 Selected from the following:

[0657]

[0658] in

[0659] R 2a R 2a’ Independently selected from H, T, or F;

[0660] R 2b It is independently selected from T, F, -OH, C1-C4 alkyl, halo-C1-C4 alkyl, -NH2, -CN, CT3 or C1-C4 alkoxy;

[0661] R 2c R 2c’ It is independently selected from T, H, F, OH, OCH3, CT3, or CH3;

[0662] R 2dSelected from T, H, F or -OH;

[0663] R 2e Selected from T, H, OH, CH3, CT3, or F;

[0664] Z can be N, NH, N (C1-C4 alkyl), N (halogenated C1-C4 alkyl), O, or S independently;

[0665] Z 1 It can be N, NH, O, or S independently;

[0666] p is 0, 1, or 2;

[0667] m is 0 or 1;

[0668] When the valence is allowed, then It is a combination of single and double bonds;

[0669] Fluorine is 19 F;

[0670] The C1-C4 alkyl, halo-C1-C4 alkyl, or C1-C4 alkoxy group optionally contains one or more T atoms, and

[0671] * indicates the location of the bond.

[0672] Preferably, the detectable labeled compound of formula (III-H) or its stereoisomers, racemic mixtures, pharmaceutically acceptable salts, hydrates, or solvates contain one, two, or three Ts. Preferably, the detectable labeled compound of formula (III-Ha) or its stereoisomers, racemic mixtures, pharmaceutically acceptable salts, hydrates, or solvates contain one T. More preferably, the detectable labeled compound of formula (III-Ha) contains two Ts. Even more preferably, the detectable labeled compound of formula (III-Ha) contains three Ts.

[0673] Preferably, the detectable labeled compound of formula (III-H) or its stereoisomers, racemic mixtures, pharmaceutically acceptable salts, hydrates, or solvates are compounds of formula (III-Ha).

[0674]

[0675] Or its stereoisomers, racemic mixtures, pharmaceutically acceptable salts, hydrates, or solvates.

[0676] in

[0677] R 1 -CN; or halogen; or C1-C4 alkyl; or C1-C4 alkoxy; or -N(C1-C4 alkyl)2; or -NH(C1-C4 alkyl); or H; or

[0678] R 1 It is a -NH-C3-C6 cycloalkyl, C3-C6 cycloalkyl or heterocyclic group, each of which is optionally substituted with at least one halogen;

[0679] R 1 Selected from:

[0680]

[0681] R 2 It is an aryl, 5-membered, or 6-membered heteroaryl group, wherein R 2 Selected from the following, where R 2 It may be replaced by at least one T.

[0682]

[0683] and

[0684] in

[0685] R 2a R 2a’ Independently selected from H, T, or F;

[0686] R 2b Independently selected from T, F, -OH, C1-C4 alkyl, halo-C1-C4 alkyl, -NH2, -CN, CT3 or C1-C4 alkoxy, wherein the C1-C4 alkyl, halo-C1-C4 alkyl or C1-C4 alkoxy optionally includes one or more T;

[0687] R 2c R 2c’ It is independently selected from T, H, F, OH, OCH, CT3, or CH3;

[0688] R 2d Selected from T, H, F or -OH;

[0689] R 2e Selected from T, H, OH, CH3, CT3, or F;

[0690] Z can be N, NH, N (C1-C4 alkyl), N (halogenated C1-C4 alkyl), O, or S independently;

[0691] Z 1 It can be N, NH, O, or S independently;

[0692] p is 0, 1, or 2;

[0693] m is 0 or 1;

[0694] When valence is allowed It is a combination of single and double bonds.

[0695] T is 3 H (tritium);

[0696] n is 0-3;

[0697] The condition is that the compound of formula (I-Ha) contains at least one T;

[0698] Fluorine is 19 F; and

[0699] * indicates the location of the bond.

[0700] Preferably, the detectable labeled compound of formula (III-Ha) or its stereoisomers, racemic mixtures, pharmaceutically acceptable salts, hydrates, or solvates contain one, two, or three Ts. Preferably, n is 1.

[0701] Preferably, the detectable labeled compound of formula (III-Ha) or its stereoisomers, racemic mixtures, pharmaceutically acceptable salts, hydrates, or solvates comprises one T. More preferably, the detectable labeled compound of formula (III-Ha) or its stereoisomers, racemic mixtures, pharmaceutically acceptable salts, hydrates, or solvates comprises two Ts. Even more preferably, the detectable labeled compound of formula (III-Ha) or its stereoisomers, racemic mixtures, pharmaceutically acceptable salts, hydrates, or solvates comprises three Ts.

[0702] In another embodiment, the present invention provides a detectable labeled compound of formula (III-H) or (111-Ha) as disclosed above, or a stereoisomer thereof, a racemic mixture, a pharmaceutically acceptable salt, a hydrate, or a solvate thereof, wherein R2 is an aryl or a 5- or 6-membered heteroaryl group selected from...

[0703]

[0704] Where R 2a R 2a’ R 2b R 2c R 2c’ R 2 d, R 2e As defined above, R... z It is selected from T, H, C1-C4 alkyl, CT3 or halogenated C1-C4 alkyl; wherein the C1-C4 alkyl and halogenated C1-C4 alkyl optionally include one or more T.

[0705] Preferably, R 2 Selected from the following:

[0706]

[0707] Where R 2a R 2a’ R 2b R 2c R 2c’ R 2d R 2e R z p is as defined above.

[0708] More preferably, R 2 Selected from the following:

[0709]

[0710] Where R 2a R 2a’ R 2b R 2c R 2c’ R 2e R z p is as defined above.

[0711] Preferably, R 2 for Where R z It contains at least one T.

[0712] More preferably, R 2 is

[0713] Preferred detectable labeled compounds of formula (III-H) or (III-Ha), their pharmaceutically acceptable salts, hydrates, or solvates are

[0714]

[0715] Where T refers to 3 H (tritium). Preferred F refers to... 19 F.

[0716] In a preferred embodiment, the present invention provides detectable labeled compounds of formula (III-H) or (III-Ha) or their stereoisomers, racemic mixtures, pharmaceutically acceptable salts, hydrates, or solvates, wherein... 3 H-tritium (“T”) can be 2 H-deuterium (“D”) is used as a substitute.

[0717] Preferably, the detectable label of a compound of formula (I) or its subforms (e.g., (IIa), (IIb), (IIIa), (IIIb), (IIIc), (III-F), (III-H)) or its stereoisomers, racemic mixtures, pharmaceutically acceptable salts, hydrates, or solvates contains a detectable label, preferably a radioactive isotope, particularly selected from... 18 F, 2 H and 3 H.

[0718] Compounds of the present invention, or detectably labeled compounds thereof, stereoisomers, racemic mixtures, pharmaceutically acceptable salts, hydrates, or solvates, and their precursors, can be detectably labeled. There is no particular limitation on the type of label, but rather it depends on the chosen detection method. Examples of possible labels include isotopes, such as radionuclides, positron emitters, and gamma emitters. Regarding detectably labeled compounds of the present invention, or their stereoisomers, racemic mixtures, pharmaceutically acceptable salts, hydrates, or solvates, and their precursors, including radioactive isotopes, positron emitters, or gamma emitters, it should be understood that the radioactive isotopes, positron emitters, or gamma emitters are present in amounts different from the naturally occurring amounts of the corresponding radioactive isotopes, positron emitters, or gamma emitters. Furthermore, the amounts used should allow for detection by the chosen detection method.

[0719] Examples of suitable isotopes such as radionuclides, positron emitters, and gamma emitters include 2 H, 3 H, 18 F, 11 C 13 N and 15 O, preferred 2 H, 3 H, 11 C 13 N、 15 O and 18 F, more preferred 2 H, 3 H and 18 F, or even better 3 H and 18 F.

[0720] 18 F-labeled compounds are particularly suitable for imaging applications, such as PET. These include compounds with natural... 19 The corresponding compounds of F isotopes are also of particular interest because they can be found in their... 18 F-analytes are used as analytical standards and references during manufacturing, quality control, release, and clinical use.

[0721] In addition, isotopes such as deuterium (i.e. 2 H) substitution can provide certain diagnostic and therapeutic advantages due to greater metabolic stability achieved by reducing, for example, defluorination, increasing in vivo half-life, or reducing dosage requirements, while maintaining or improving the efficacy of the original compound.

[0722] The isotopic variations of the compounds of the present invention or their detectable labeled compounds, stereoisomers, racemic mixtures, pharmaceutically acceptable salts, hydrates or solvates and their precursors can generally be prepared by conventional methods, such as by exemplary methods or by the preparation methods described in the examples and preparation examples below, using appropriate isotopic variations of suitable reagents, which are commercially available or prepared by known synthetic techniques.

[0723] The compounds of the present invention, or their detectable-labeled compounds, stereoisomers, racemic mixtures, pharmaceutically acceptable salts, hydrates, or solvates, and their precursors, can be incorporated into radionuclides, positron emitters, and gamma emitters using methods commonly used in the field of organic synthesis. Typically, when preparing the desired compounds of the present invention, or their detectable-labeled compounds, stereoisomers, racemic mixtures, pharmaceutically acceptable salts, hydrates, or solvates, and their precursors, they can be introduced by using appropriately labeled starting materials. Exemplary methods for introducing detectable labels are described, for example, in US2012 / 0302755.

[0724] There are no particular limitations on the location where the detectable marker binds to the compounds and precursors of the present invention.

[0725] For example, radioactive nuclides, positron emitters, and gamma emitters can be attached to any position where corresponding non-emitting atoms can also be attached. 18 F can be attached to any suitable location for F attachment. This also applies to other radionuclides, positron emitters, and gamma emitters. Due to ease of synthesis, it is preferred to attach F at R. 1 Connection 18 F. 3 H can be attached to any available location. It is preferred to be attached to a pyridine ring. If 2 If H is used as a detectable marker, it can be attached to any available location. Preferably, it is attached to a pyridine ring.

[0726] In another embodiment, the invention also relates to compounds of formula (IV-F), i.e., precursors of compounds of formula (III-F).

[0727]

[0728] in

[0729] R 3The following are replaced by leaving groups (LG):

[0730] R 4 It is an aryl, 5-membered, or 6-membered heteroaryl group, wherein R 4 R selected from compounds of formula (III-F) disclosed above 2 Same list.

[0731] Preferably, the leaving group (LG) is halogen or C. 1-4 Alkyl sulfonates, C1-C4 alkylammonium, nitro or C 6-10 Aryl sulfonates, wherein C 6-10 The aryl group may optionally be substituted with -CH3 or -NO2. More preferably, the leaving group (LG) is bromine, chlorine, iodine, or C. 1-4 Alkyl sulfonates or C 6-10 Aryl sulfonates, wherein C 6-10 The aryl group may optionally be substituted with -CH3 or -NO2. Even more preferably, the leaving group (LG) is a methanesulfonate, toluenesulfonate, or nitrobenzenesulfonate. Even more preferably, the leaving group (LG) is a methanesulfonate or nitrobenzenesulfonate. Preferably, the leaving group (LG) is a methanesulfonate.

[0732] Preferably, R 4 for

[0733] More preferably, R 4 for

[0734] Even more preferably, R 4 for

[0735] Preferably, R 4 Optional 18 F is replaced.

[0736] The preferred compounds are:

[0737]

[0738] In another embodiment, the invention also relates to compounds of formula (IV-H) or their stereoisomers, racemic mixtures, pharmaceutically acceptable salts, hydrates, or solvates, i.e., precursors of compounds of formula (III-H) or their stereoisomers, racemic mixtures, pharmaceutically acceptable salts, hydrates, or solvates.

[0739]

[0740] in

[0741] R 5 R selected from compounds of formula (III-H) disclosed above 1 Same list, and preferred from

[0742]

[0743] R 6 It is an aryl, 5-membered, or 6-membered heteroaryl group, wherein R 6 Selected from the following:

[0744]

[0745] in

[0746] R 2a R 2a’ Independently selected from H, X, or F;

[0747] R 2b Independently selected from X, F, -OH, C1-C4 alkyl, halo-C1-C4 alkyl, -NH2, -CN or C1-C4 alkoxy, wherein the C1-C4 alkyl, halo-C1-C4 alkyl or C1-C4 alkoxy optionally includes one or more X;

[0748] R 2c R 2c’ It is independently selected from X, H, F, OH, OCH3 or CH3;

[0749] R 2d Selected from X, H, F, or -OH;

[0750] R 2e Selected from X, H, OH, CH3, or F;

[0751] z can be N, NH, N (C1-C4 alkyl), N (halogenated C1-C4 alkyl), O, or S independently;

[0752] Z 1 It can be N, NH, O, or S independently;

[0753] p is 0, 1, or 2;

[0754] m is 0 or 1;

[0755] When valence is allowed It is a combination of single and double bonds;

[0756] * indicates the location of the bond.

[0757] Fluorine is 19 F;

[0758] X is bromine, chlorine, or iodine; and

[0759] Where R 6 It contains at least one X.

[0760] In another embodiment, in a compound of formula (IV-H) or its stereoisomers, racemic mixtures, pharmaceutically acceptable salts, hydrates, or solvates, R 6 Preferably, the aryl group or 6-heteroaryl group is optionally substituted with one or more X groups, selected from:

[0761]

[0762] Where R 2a R 2a’ R 2b R 2c R 2c’ R 2d R 2e And p as defined above; when valence allows, It is a combination of single and double bonds; fluorine is 19 F; and * indicates the location of the bond.

[0763] Preferably, R 6 for More preferably, R 6 for

[0764] Even more preferably, the compound of formula (IV-H) is:

[0765]

[0766] Or its detectable labeled compounds, stereoisomers, racemic mixtures, pharmaceutically acceptable salts, hydrates, or solvates.

[0767] X is selected from bromine, chlorine, and iodine.

[0768] Preferably, X is bromine.

[0769] The preferred compounds are:

[0770]

[0771] Or a detectable labeled compound, a pharmaceutically acceptable salt, hydrate, or solvate.

[0772] In another embodiment, the invention also relates to compounds of formula (IV-J) or their stereoisomers, racemic mixtures, pharmaceutically acceptable salts, hydrates, or solvates, which are precursors of compounds of formula (III-H) or their stereoisomers, racemic mixtures, pharmaceutically acceptable salts, hydrates, or solvates.

[0773]

[0774] in

[0775] R 7 R selected from compounds of formula (III-H) disclosed above 1 Same list, and preferred from

[0776]

[0777] R 8 Selected from the following:

[0778]

[0779] in

[0780] R 2a R 2a’ Independently selected from H or F;

[0781] R 2b Independently selected from F, -OH, C1-C4 alkyl, halo-C1-C4 alkyl, -NH2, -CN or C1-C4 alkoxy;

[0782] p is 0, 1, or 2;

[0783] R z Selected from H, C1-C4 alkyl and halo-C1-C4 alkyl,

[0784] When valence is allowed It is a combination of single and double bonds;

[0785] Fluorine is 19 F; and

[0786] * indicates the location of the bond.

[0787] Preferably, R z For H.

[0788] In another embodiment of a compound of formula (IV-J) or its stereoisomers, racemic mixtures, pharmaceutically acceptable salts, hydrates, or solvates, R 8 Preferred from

[0789]

[0790] Where R 2a R 2a’ R 2b p is as defined above;

[0791] More preferably, R 8 Selected from:

[0792] The preferred compounds are:

[0793]

[0794] Or a detectable labeled compound, a pharmaceutically acceptable salt, hydrate, or solvate.

[0795] Synthesis methods for detectable labeled compounds

[0796] The present invention also relates to methods for preparing compounds of formula (I) or its sub-forms (e.g., (IIa), (IIb), (IIIa), (IIIb), (IIIc), (III-F), (III-H)) or their detectable labeled compounds, stereoisomers, racemic mixtures, pharmaceutically acceptable salts, hydrates or solvates, and particularly methods comprising detectable labeled compounds of formula (III-F) or (III-H) or their stereoisomers, racemic mixtures, pharmaceutically acceptable salts, hydrates or solvates.

[0797] In one embodiment, the present invention relates to a method for preparing compounds of formula (III-F) or their stereoisomers, racemic mixtures, pharmaceutically acceptable salts, hydrates, or solvates, wherein the method involves using a radioisotope 18 F is used to radiolabel compounds of formula (IV-F) for this purpose.

[0798] 18 F-fluorinating agent

[0799]

[0800] Where R 1 R 2 R 3 and R 4 As defined in this article.

[0801] For 18 Suitable solvents for F-fluorination include DMF, DMSO, acetonitrile, DMA, or mixtures thereof, preferably acetonitrile or DMSO.

[0802] For 18 Suitable reagents for F-fluorination are selected from K 18 F, Rb 18 F, Cs 18 F, Na 18 F, 18 F's four (C) 1-6 Alkyl ammonium salts, kryptofix

[222] 18 F and tetrabutylammonium fluoride [ 18 F).

[0803] In one embodiment, the present invention relates to a method for preparing compounds of the formula (III-H) or their stereoisomers, racemic mixtures, pharmaceutically acceptable salts, hydrates, or solvates, by means of a radioisotope. 3 H is used to radiolabel compounds of formula (IV-H) for this purpose.

[0804] 3 H radiolabeled reagent

[0805]

[0806] Where R 1 R 2 R 5 and R 6 As defined in this article, and

[0807] T is 3 H (tritium),

[0808] n is 0-3, preferably n is 1 or 2, more preferably n is 1;

[0809] The condition is that the compound of formula (III-Ha) contains at least one T.

[0810] Fluorine is 19 F,

[0811] X is bromine, chlorine, iodine or H, preferably bromine.

[0812] 3 The H radiolabeling reagent can be tritium gas. This method can be carried out in the presence of a catalyst such as palladium / carbon (Pd / C), a solvent such as dimethylformamide (DMF), and a base such as N,N-diisopropylethylamine (DIEA).

[0813] In a preferred embodiment, F (fluorine) is... 19 F.

[0814] In one embodiment, the present invention relates to a method for preparing compounds of formula (III-H) or their stereoisomers, racemic mixtures, pharmaceutically acceptable salts, hydrates, or solvates, by radiolabeling compounds of formula (IV-J) with a CT3 radiolabeling reagent, wherein T is... 3 H.

[0815]

[0816] CT3 radiolabeling reagent can be ICT3 (containing 3 (Iodomethane derivatives of H). This method can be carried out in the presence of a solvent such as dimethylformamide (DMF) and a base such as cesium carbonate or sodium hydride.

[0817] Diagnostic Compositions

[0818] The compounds of the present invention, or detectable and labelable compounds thereof, stereoisomers, racemic mixtures, pharmaceutically acceptable salts, hydrates, or solvates, are particularly suitable for imaging α-synuclein aggregates (including, but not limited to, Lewy bodies and / or Lewy neurites). Regarding α-synuclein aggregates, the compounds are particularly suitable for binding different types of α-synuclein aggregates, including, but not limited to, Lewy bodies and / or Lewy neurites. Imaging can be performed in mammals, preferably in humans. Imaging is preferably in vitro, ex vivo, or in vivo. More preferably, imaging is in vivo; even more preferably, imaging is brain imaging. Imaging can also be eye / retinal imaging. The compounds of the present invention, or detectable and labelable compounds thereof, stereoisomers, racemic mixtures, pharmaceutically acceptable salts, hydrates, or solvates, are particularly suitable for diagnostic purposes.

[0819] Diagnosis can be performed on mammals, preferably humans. The tissues of interest for diagnosis can be the brain, central nervous system tissues, eye tissues (e.g., retinal tissues), or other tissues or fluids, such as cerebrospinal fluid (CSF). The preferred tissue is brain tissue.

[0820] Due to their design and binding characteristics, the compounds of the present invention, or detectable and labeled compounds thereof, stereoisomers, racemic mixtures, pharmaceutically acceptable salts, hydrates, or solvates, are suitable for the diagnosis of diseases, conditions, and abnormalities associated with α-synuclein aggregates (including, but not limited to, Lewy bodies and / or Lewy neurites). The compounds of the present invention, or detectable and labeled compounds thereof, stereoisomers, racemic mixtures, pharmaceutically acceptable salts, hydrates, or solvates, are particularly suitable for positron emission tomography (PET) of α-synuclein aggregates (including, but not limited to, Lewy bodies and / or Lewy neurites). Diseases involving α-synuclein aggregates are generally classified as synucleinopathies (or α-synucleinopathies). The compounds of this invention, or their detectable labeled compounds, stereoisomers, racemic mixtures, pharmaceutically acceptable salts, hydrates, or solvates, are suitable for diagnosing diseases, symptoms, or abnormalities, including but not limited to Parkinson's disease (sporadic, familial with α-synuclein mutations or familial with mutations other than α-synuclein, simple autonomic failure, and Lewy body dysphagia), SNCA repeat carriers, Lewy body dementia (“simple” Lewy body dementia), Alzheimer's disease, sporadic Alzheimer's disease, familial Alzheimer's disease with APP mutations, familial Alzheimer's disease with PS-1, PS-2, or other mutations, familial British dementia, Lewy body variants of Alzheimer's disease, and normal aging in Down syndrome. Synucleinic diseases involving α-synuclein aggregates of neurons and glial cells include, but are not limited to, multiple system atrophy (MSA) (Shy-Drager syndrome, striatal substantia nigra, and olivary pontocerebellar atrophy).Other diseases that may involve α-synuclein immunoreactive lesions include traumatic brain injury, chronic traumatic encephalopathy, tau disorders (Pick's disease, frontotemporal dementia, progressive supranuclear palsy, corticobasal degeneration, and Niemann-Pick type C1 disease), motor neuron disease, amyotrophic lateral sclerosis (sporadic, familial, and Guam ALS-dementia syndrome), axillary dystrophy, type I brain iron accumulation neurodegeneration (Has-Schwarz syndrome), prion diseases, ataxia-telangiectasia, idiopathic orofacial motor disorders, subacute sclerosing panencephalitis, Gaucher disease, and other lysosomal storage disorders (including Kufor-Rakeb syndrome and Sanfilippo syndrome) and rapid eye movement (REM) sleep behavior disorder (Jellinger, MovDisord 2003, 18 Supplement 6, S2-12; Galvin et al., JAMA Neurology). 2001, 58(2), 186-190; Kovari et al., Acta Neuropathol. 2007, 114(3), 295-8; Saito et al., J Neuropathol ExpNeurol. 2004, 63(4), 323-328; McKee et al., Brain, 2013, 136(Pt 1), 43-64; Puschmann et al., Parkinsonism Relat Disord 2012, 18S1, S24-S27; Usenovic et al., J Neurosci. 2012, 32(12), 4240-4246; Winder-Rhodes et al., Mov Disord. 2012, 27(2), 312-315; Ferman et al., J Int Neuropsychol Soc. 2002, 8(7), 907-914). Preferably, the compounds of the present invention are suitable for diagnosing Parkinson's disease, multiple system atrophy, Lewy body dementia, Parkinson's disease dementia, SNCA repeat carriers or Alzheimer's disease, more preferably Parkinson's disease (PD).

[0821] In methods for diagnosing diseases, conditions, or abnormalities (e.g., Parkinson's disease or susceptibility) associated with α-synuclein aggregates (including but not limited to Lewy bodies and / or Lewy neurites) in individuals, the method includes the following steps:

[0822] a) Administer to an individual a diagnostically effective amount of the compound of the present invention or a detectable labeled compound thereof, stereoisomer, racemic mixture, pharmaceutically acceptable salt, hydrate or solvate;

[0823] b) Allows the compounds of the present invention or their detectable labeled compounds, stereoisomers, racemic mixtures, pharmaceutically acceptable salts, hydrates, or solvates to distribute into the tissue of interest (e.g., brain or other tissues or body fluids, such as cerebrospinal fluid (CSF)); and

[0824] c) Imaging the tissue of interest, wherein increased binding of the compound of the present invention or its detectable labeled compound, stereoisomer, racemic mixture, pharmaceutically acceptable salt, hydrate or solvate to the tissue of interest, compared to normal control binding levels, indicates that the individual has a disease, condition or abnormality associated with α-synuclein aggregates or is at risk of developing a disease, condition or abnormality associated with α-synuclein aggregates, including but not limited to Lewy bodies and / or Lewy neurites.

[0825] The compounds of the present invention, or detectable and labelable compounds thereof, stereoisomers, racemic mixtures, pharmaceutically acceptable salts, hydrates, or solvates, can be used to image α-synuclein aggregates (including, but not limited to, Lewy bodies and / or Lewy neurites) in any sample from a patient suspected of containing α-synuclein aggregates (including, but not limited to, Lewy bodies and / or Lewy neurites) or in a specific body site or region. These compounds are capable of crossing the blood-brain barrier. Therefore, they are particularly suitable for imaging α-synuclein aggregates, including, but not limited to, Lewy bodies and / or Lewy neurites, in the brain or peripheral organs such as the intestines and in body fluids such as cerebrospinal fluid (CSF).

[0826] In diagnostic applications, compounds of the present invention or their detectable-labeled compounds, stereoisomers, racemic mixtures, pharmaceutically acceptable salts, hydrates, or solvates, preferably of formula (I) or its subforms (e.g., (IIa), (IIb), (IIIa), (IIIb), (IIIc), (III-F), (III-H)), are preferably administered in the form of diagnostic compositions comprising compounds of the present invention or their detectable-labeled compounds, stereoisomers, racemic mixtures, pharmaceutically acceptable salts, hydrates, or solvates. In this invention, a "diagnostic composition" is defined as a composition comprising one or more compounds of the present invention or their detectable-labeled compounds, stereoisomers, racemic mixtures, pharmaceutically acceptable salts, hydrates, or solvates, in a form suitable for administration to a patient, such as a mammal, such as a human, and suitable for diagnosing said specific disease, condition, or abnormality. Preferably, the diagnostic composition further comprises physiologically acceptable excipients, diluents, or adjuvants. Administration is preferably carried out as defined below. More preferably, by injection of the composition as an aqueous solution. Such compositions may optionally contain additional components, such as buffers; pharmaceutically acceptable solubilizers (e.g., cyclodextrins or surfactants, such as prolind, Tween, or phospholipids); and pharmaceutically acceptable stabilizers or antioxidants (e.g., ascorbic acid, gentianic acid, or para-aminobenzoic acid). The dosage of the compounds of the present invention or their detectable-labeled compounds, stereoisomers, racemic mixtures, pharmaceutically acceptable salts, hydrates, or solvates may vary depending on the specific compound administered, the patient's weight, and other variables readily apparent to a skilled clinician in the art.

[0827] While it is possible that the compounds of the present invention or their detectable-labeled compounds, stereoisomers, racemic mixtures, pharmaceutically acceptable salts, hydrates, or solvates may be administered alone, they are preferably formulated into diagnostic compositions according to standard pharmaceutical practice. Therefore, the present invention also provides diagnostic compositions comprising a diagnostically effective amount of the compounds of the present invention or their detectable-labeled compounds, stereoisomers, racemic mixtures, pharmaceutically acceptable salts, hydrates, or solvates, and optionally at least one pharmaceutically acceptable excipient, carrier, diluent, or adjuvant.

[0828] Pharmaceutically acceptable excipients are well-known in the pharmaceutical field and are described, for example, in Remington's Pharmaceutical Sciences, 15th edition, Mack Publishing Co., New Jersey (1975). Pharmaceutical excipients can be selected based on the intended route of administration and standard pharmaceutical practice. Excipients must be acceptable in the sense that they are harmless to the recipient.

[0829] Pharmaceutically useful excipients, carriers, adjuvants, and diluents that can be used in formulations of the diagnostic compositions of the present invention may include, for example, solvents such as monohydric alcohols (e.g., ethanol, isopropanol) and polyhydric alcohols (e.g., diols) and edible oils (such as soybean oil, coconut oil, olive oil, safflower oil, cottonseed oil), oil esters such as ethyl oleate, isopropyl myristate, binders, adjuvants, solubilizers, thickeners, stabilizers, disintegrants, flow aids, lubricants, buffers, emulsifiers, wetting agents, suspending agents, sweeteners, colorants, flavoring agents, coatings, preservatives, antioxidants, processing agents, drug delivery modifiers, and enhancers, such as calcium phosphate, magnesium stearate, talc, monosaccharides, disaccharides, starch, gelatin, cellulose, methylcellulose, sodium carboxymethylcellulose, dextran, hydroxypropyl-β-cyclodextrin, polyvinylpyrrolidone, low-melting-point waxes, and ion exchange resins.

[0830] The compounds of the present invention, preferably of formula (I) or its sub-forms (e.g., (IIa), (IIb), (IIIa), (IIIb), (IIIc), (III-F), (III-H)), or their detectable labeled compounds, stereoisomers, racemic mixtures, pharmaceutically acceptable salts, hydrates, or solvates, may be administered (delivered) via routes including, but not limited to, one or more of the following: intravenous, gastrointestinal, spinal, peritoneal, intramuscular, oral (e.g., tablets, capsules, or ingestible solutions), topical, mucosal (e.g., nasal sprays or aerosols for inhalation), nasal, parenteral (e.g., via injectable form), intrauterine, intraocular, intradermal, intracranial, intratracheal, intravaginal, intravenous, intracerebral, subcutaneous, ocular (including vitreous or anterior chamber), transdermal, rectal, sublingual, epidural, and sublingual. Preferably, the compounds of the present invention, or their detectable labeled compounds, stereoisomers, racemic mixtures, pharmaceutically acceptable salts, hydrates, or solvates, are administered (delivered) intravenously.

[0831] For example, the compound can be administered orally in the form of tablets, capsules, ovules, elixirs, solutions or suspensions, and may contain flavoring agents or coloring agents for immediate release, delayed release, improved release, sustained release, pulsatile release or controlled release applications.

[0832] Tablets may contain excipients such as microcrystalline cellulose, lactose, sodium citrate, calcium carbonate, calcium hydrogen phosphate, and glycine; disintegrants such as starch (preferably corn, potato, or cassava starch), sodium glycolate starch, croscarmellose sodium, and certain complex silicates; and granulation binders such as polyvinylpyrrolidone, hydroxypropyl methylcellulose (HPMC), hydroxypropyl cellulose (HPC), sucrose, gelatin, and gum arabic. Additionally, lubricants such as magnesium stearate, stearic acid, glyceryl behenate, and talc may be included. Similar types of solid compositions may also be used as fillers in gelatin capsules. In this regard, preferred excipients include starch, cellulose, milk sugar (lactose), or high molecular weight polyethylene glycol. For use in aqueous suspensions and / or elixirs, the active agent may be combined with various sweeteners or flavoring agents, colorants or dyes, emulsifiers and / or suspending agents, and diluents such as water, ethanol, propylene glycol, and glycerin, and combinations thereof.

[0833] Preferably, in diagnostic applications, the compounds of the present invention or their detectable-labeled compounds, stereoisomers, racemic mixtures, pharmaceutically acceptable salts, hydrates, or solvates are administered parenterally. If the compounds of the present invention or their detectable-labeled compounds, stereoisomers, racemic mixtures, pharmaceutically acceptable salts, hydrates, or solvates are administered parenterally, examples of such administration include one or more of the following: intravenous, intra-arterial, intraperitoneal, intrathecal, intraventricular, intraurethral, ​​intrasternal, intracranial, intramuscular, or subcutaneous administration; and / or administration using infusion techniques. For parenteral administration, the compounds are preferably used in the form of a sterile aqueous solution, which may contain other substances, such as sufficient salts or glucose, to make the solution isotonic with blood. If necessary, the aqueous solution should be appropriately buffered (preferably pH 3-9). The preparation of suitable parenteral formulations under sterile conditions can be readily accomplished using standard pharmaceutical techniques well known to those skilled in the art.

[0834] As shown, the compounds of the present invention or their detectable-labeled compounds, stereoisomers, racemic mixtures, pharmaceutically acceptable salts, hydrates, or solvates can be administered intranasally or by inhalation and conveniently delivered from a pressurized container, pump, nebulizer, or sprayer in the form of a dry powder inhaler or aerosol spray, wherein a suitable propellant is used, such as dichlorodifluoromethane, trichlorofluoromethane, dichlorotetrafluoroethane, hydrofluoroalkane such as 1,1,1,2-tetrafluoroethane (HFA134AT) or 1,1,1,2,3,3,3-heptafluoropropane (HFA227EA), carbon dioxide, or other suitable gas. In the case of pressurized aerosols, the dosage unit can be determined by providing a valve for delivering a metered amount. The pressurized container, pump, nebulizer, or sprayer may contain a solution or suspension of the active compound, for example using a mixture of ethanol and propellant as a solvent, and may additionally contain a lubricant, such as sorbitan trioleate. Capsules and cartridges for inhalers or blowpipes (e.g., made of gelatin) can be formulated into powder mixtures containing compounds and suitable powder matrices such as lactose or starch.

[0835] Alternatively, the compounds of the present invention or their detectable labeled compounds, stereoisomers, racemic mixtures, pharmaceutically acceptable salts, hydrates, or solvates may be administered in the form of suppositories or vaginal suppositories, or topically in the form of gels, hydrogels, lotions, solutions, creams, ointments, or powders. The compounds of the present invention or their detectable labeled compounds, stereoisomers, racemic mixtures, pharmaceutically acceptable salts, hydrates, or solvates may also be administered through the skin or transdermally, for example, by using skin patches.

[0836] They can also be administered via the lungs or rectum. They can also be administered via the eyes. For ophthalmic use, the compounds can be formulated as micronized suspensions in isotonic, pH-adjusted sterile saline, or preferably as solutions in isotonic, pH-adjusted sterile saline, optionally in combination with a preservative such as benzalkonium chloride. Alternatively, they can be formulated as ointments, such as petrolatum.

[0837] For topical application to the skin, the compounds of the present invention or their detectable-labeled compounds, stereoisomers, racemic mixtures, pharmaceutically acceptable salts, hydrates, or solvates can be formulated into suitable ointments comprising an active compound suspended or dissolved in, for example, a mixture with one or more of the following: mineral oil, petrolatum, white petrolatum, propylene glycol, emulsifying wax, and water. Alternatively, they can be formulated into suitable lotions or creams suspended or dissolved in, for example, a mixture with one or more of the following: mineral oil, stearyl sorbitan, polyethylene glycol, liquid paraffin, polysorbate 60, cetyl ester wax, cetearyl alcohol, 2-octyldodecanool, benzyl alcohol, and water.

[0838] Typically, clinicians will determine the actual dose best suited for the individual. The specific dose level and frequency of administration for any particular individual can vary and will depend on a variety of factors, including the activity of the particular compound used, the metabolic stability and duration of action of the compound, age, weight, general health condition, sex, diet, route and time of administration, excretion rate, drug combination, severity of the specific condition, and the individual undergoing diagnosis.

[0839] The diagnostic compositions of the present invention can be produced in a manner known to those skilled in the art, for example as described in Remington's Pharmaceutical Sciences, 15th edition, Mack Publishing Co., New Jersey (1975).

[0840] The compounds of the present invention, or their detectable and labeled compounds, stereoisomers, racemic mixtures, pharmaceutically acceptable salts, hydrates, or solvates, are used as in vitro analytical references or in vitro screening tools. They are also used in in vivo diagnostic methods.

[0841] The compounds of the present invention or their detectable labeled compounds, stereoisomers, racemic mixtures, pharmaceutically acceptable salts, hydrates, or solvates can also be provided in the form of mixtures comprising the compounds of the present invention or their detectable labeled compounds, stereoisomers, racemic mixtures, pharmaceutically acceptable salts, hydrates, or solvates, and at least one compound selected from imaging agents, pharmaceutically acceptable excipients, carriers, diluents, or adjuvants of non-inventive compounds. Imaging agents of non-inventive compounds are preferably present in a diagnostically effective amount. More preferably, the imaging agents of non-inventive compounds are Aβ or Tau imaging agents.

[0842] The diagnosis of a patient's susceptibility to diseases, conditions, or abnormalities associated with α-synuclein aggregates (including but not limited to Lewy bodies and / or Lewy neurites) can be achieved by detecting the specific binding of the compound of the present invention or its detectable labeled compounds, stereoisomers, racemic mixtures, pharmaceutically acceptable salts, hydrates, or solvates to α-synuclein aggregates (including but not limited to Lewy bodies and / or Lewy neurites) in a sample or a specific body site or region, comprising the following steps:

[0843] (a) Exposing a sample or specific body part or region suspected of containing α-synuclein aggregates (including but not limited to Lewy bodies and / or Lewy neurites) to the compound of the present invention or its detectable labeled compound, stereoisomer, racemic mixture, pharmaceutically acceptable salt, hydrate, or solvate thereof, which binds to α-synuclein aggregates (including but not limited to Lewy bodies and / or Lewy neurites),

[0844] (b) Allows the compounds of the present invention or their detectable labeled compounds, stereoisomers, racemic mixtures, pharmaceutically acceptable salts, hydrates or solvates to bind to α-synuclein aggregates (including but not limited to Lewy bodies and / or Lewy neurites) to form compound / (α-synuclein aggregates, including but not limited to Lewy bodies or Lewy neurites) complexes (hereinafter, "compound / (α-synuclein aggregates (including but not limited to Lewy bodies and / or Lewy neurites)) complexes", abbreviated as "compound / protein aggregate complexes").

[0845] (c) Detection of the formation of compound / protein aggregate complexes.

[0846] (d) Optionally establish a correlation between the presence or absence of the compound / protein aggregate complex and the presence or absence of α-synuclein aggregates (including, but not limited to, Lewy bodies and / or Lewy neurites) in the sample or a specific body part or region; and

[0847] (e) Optionally compare the amount of the compound / protein aggregate complex with normal control values, wherein an increase in the amount of the compound / protein aggregate complex compared with normal control values ​​may indicate that the patient has a disease, condition or abnormality associated with α-synuclein aggregates (including but not limited to Lewy bodies and / or Lewy neurites) or is at risk of developing them.

[0848] The compounds of the present invention or their detectable-labeled compounds, stereoisomers, racemic mixtures, pharmaceutically acceptable salts, hydrates, or solvates can be brought into contact with samples or specific body sites or regions suspected of containing α-synuclein aggregates (including, but not limited to, Lewy bodies and / or Lewy neurites) by suitable methods. In in vitro methods, the compounds of the present invention or their detectable-labeled compounds, stereoisomers, racemic mixtures, pharmaceutically acceptable salts, hydrates, or solvates can be simply mixed with a liquid sample. In in vivo testing, the compounds of the present invention or their detectable-labeled compounds, their stereoisomers, racemic mixtures, pharmaceutically acceptable salts, hydrates, or solvates can typically be administered to a patient by any suitable means. These routes of administration include, but are not limited to, one or more of the following: oral (e.g., as tablets, capsules, or as an ingestible solution), topical, mucosal (e.g., as a nasal spray or aerosol for inhalation), nasal, parenteral (e.g., via injectable form), gastrointestinal, intraspinal, intraperitoneal, intramuscular, intravenous, intrauterine, intraocular, intradermal, intracranial, intratracheal, intravaginal, intravenous, intracerebral, subcutaneous, ocular (including vitreous or anterior chamber), transdermal, rectal, sublingual, epidural, and sublingual. In some cases, parenteral administration may be preferred.

[0849] After a sample or a specific body site or region has been contacted with the compound of the present invention or a detectable labeled compound thereof, stereoisomer, racemic mixture, pharmaceutically acceptable salt, hydrate, or solvate, the compound is allowed to bind to α-synuclein aggregates (including, but not limited to, Lewy bodies and / or Lewy neurites). The time required for binding will depend on the type of test (e.g., in vitro or in vivo) and can be determined by those skilled in the art through routine experiments.

[0850] Compounds already bound to α-synuclein aggregates (including, but not limited to, Lewy bodies and / or Lewy neurites) can then be detected by any suitable method. The specific method chosen will depend on the selected detectable marker. Examples of possible methods include, but are not limited to, fluorescence imaging techniques or nuclear imaging techniques, such as positron emission tomography (PET), single-photon emission computed tomography (SPECT), magnetic resonance imaging (MRI), and contrast-enhanced magnetic resonance imaging (MRI). These have been described and enable the visualization of amyloid biomarkers. Fluorescence imaging techniques and / or nuclear imaging techniques can be used to monitor and / or visualize the distribution of detectable marker compounds in a sample or specific body site or region.

[0851] Optionally, a correlation can then be established between the presence or absence of the compound / protein aggregate complex and the presence or absence of α-synuclein aggregates in the sample or a specific body part or region, including but not limited to Lewy bodies and / or Lewy neurites. Finally, the amount of the compound / protein aggregate complex can be compared to normal control values ​​measured in a sample or a specific body part or region of a healthy individual, wherein an increase in the amount of the compound / protein aggregate complex compared to normal control values ​​may indicate that the patient has a disease, condition, or abnormality associated with α-synuclein aggregates or is at risk of developing such a disease, condition, or abnormality, including but not limited to Lewy bodies and / or Lewy neurites.

[0852] The present invention also relates to a method for determining the amount of α-synuclein aggregates (including but not limited to Lewy bodies and / or Lewy neurites) in tissues and / or body fluids. The method comprises the following steps:

[0853] (a) Provide a sample representing the tissue and / or body fluid being studied;

[0854] (b) Test the presence of α-synuclein aggregates (including but not limited to Lewy bodies and / or Lewy neurites) in samples using the compounds of the present invention;

[0855] (c) Determine the amount of compounds that bind to α-synuclein aggregates (including but not limited to Lewy bodies and / or Lewy neurites); and

[0856] (d) Calculate the amount of α-synuclein aggregates (including but not limited to Lewy bodies and / or Lewy neurites) in tissues and / or body fluids.

[0857] The presence of α-synuclein aggregates (including but not limited to Lewy bodies and / or Lewy neurites) in a sample can be tested using the compounds of the present invention or their detectable labeled compounds, stereoisomers, racemic mixtures, pharmaceutically acceptable salts, hydrates, or solvates. This is achieved by contacting the sample with the compounds of the present invention, allowing the compounds of the present invention to bind to the α-synuclein aggregates (including but not limited to Lewy bodies and / or Lewy neurites) to form a compound / protein aggregate complex, and detecting the formation of the compound / protein aggregate complex as described above.

[0858] Monitoring for minimal residual disease, condition, or abnormality in patients with a disease, condition, or abnormality associated with α-synuclein aggregates (including but not limited to Lewy bodies and / or Lewy neurites), said patients having been treated with a drug having the compound of the present invention or a compound with a detectable label thereof, stereoisomer, racemic mixture, pharmaceutically acceptable salt, hydrate, or solvate, can be achieved through the following steps:

[0859] (a) Expose a sample or specific body part or area suspected of containing α-synuclein aggregates (including but not limited to Lewy bodies and / or Lewy neurites) to the compound of the present invention or its detectable labeled compound, stereoisomer, racemic mixture, pharmaceutically acceptable salt, hydrate or solvate.

[0860] (b) The compound is combined with α-synuclein aggregates (including but not limited to Lewy bodies and / or Lewy neurites) to form a compound / protein aggregate complex;

[0861] (c) Detecting the formation of the compound / protein aggregate complex;

[0862] (d) Optionally, establish a correlation between the presence or absence of the compound / protein aggregate complex and the presence or absence of α-synuclein aggregates (including, but not limited to, Lewy bodies and / or Lewy neurites) in the sample or a specific body site or region; and

[0863] (e) Optionally compare the amount of the compound / protein aggregate complex with a normal control value, wherein an increase in the amount of the aggregate compared to the normal control value may indicate that the patient still has minimal residual disease, symptoms or abnormalities.

[0864] The steps (a) through (e) have been explained above.

[0865] In methods for monitoring minimal residual disease, symptoms, or abnormalities, the method may further include steps (i) to (vi) prior to step (a):

[0866] (i) Exposing a sample or a specific body part or area suspected of containing α-synuclein aggregates (including but not limited to Lewy bodies and / or Lewy neurites) to the compound of the present invention or a compound of the present invention or a detectable labeled thereof, stereoisomer, racemic mixture, pharmaceutically acceptable salt, hydrate or solvate, said compound specifically binding to α-synuclein aggregates (including but not limited to Lewy bodies and / or Lewy neurites);

[0867] (ii) To bind the compound to α-synuclein aggregates (including but not limited to Lewy bodies and / or Lewy neurites) to form a compound / (α-synuclein aggregates (including but not limited to Lewy bodies and / or Lewy neurites)) complex;

[0868] (iii) Detect the formation of the compound / (α-synuclein aggregates (including but not limited to Lewy bodies and / or Lewy neurites)) complex;

[0869] (iv) Establish a correlation between the presence or absence of the compound / (α-synuclein aggregate (including but not limited to Lewy bodies and / or Lewy neurites)) complex and the presence or absence of α-synuclein aggregate (including but not limited to Lewy bodies and / or Lewy neurites) in the sample or a specific body part or region.

[0870] (v) Optionally, compare the amount of the compound / (α-synuclein aggregate (including but not limited to Lewy bodies and / or Lewy neurites)) complex with normal control values; and

[0871] (vi) Treat patients with medication.

[0872] Optionally, the method may further include step (A) after step (d) or step (e):

[0873] (A) Compare the amount of the compound / (α-synuclein aggregate (including but not limited to Lewy bodies and / or Lewy neurites)) complex determined in step (iv) with the amount of the compound / (α-synuclein aggregate (including but not limited to Lewy bodies and / or Lewy neurites)) complex determined in step (d).

[0874] To monitor minimal residual disease, symptoms, or abnormalities over time, steps (a) through (c) and optional steps (d) and (e) of the method for monitoring minimal residual disease, symptoms, or abnormalities may be repeated once or multiple times.

[0875] In methods for monitoring minimal residual disease, symptoms, or abnormalities, the amount of the compound / protein aggregate complex can optionally be compared at various time points during treatment, such as before and after treatment begins, or at various time points after treatment begins. Changes in the amount of the compound / protein aggregate complex, particularly a decrease, can indicate that the residual disease, symptoms, or abnormalities are decreasing.

[0876] Predicting the responsiveness of patients with diseases, conditions, or abnormalities associated with α-synuclein aggregates (including but not limited to Lewy bodies and / or Lewy neurites) and who are treated with medication can be achieved through the following steps:

[0877] (a) Expose a sample or specific body part or area suspected of containing α-synuclein aggregates (including but not limited to Lewy bodies and / or Lewy neurites) to the compound of the present invention or its detectable labeled compound, stereoisomer, racemic mixture, pharmaceutically acceptable salt, hydrate or solvate.

[0878] (b) Allow the compound to bind to α-synuclein aggregates (including but not limited to Lewy bodies and / or Lewy neurites) to form a compound / protein aggregate complex;

[0879] (c) Detecting the formation of the compound / protein aggregate complex;

[0880] (d) Optionally, establish a correlation between the presence or absence of the compound / protein aggregate complex and the presence or absence of α-synuclein aggregates (including, but not limited to, Lewy bodies and / or Lewy neurites) in the sample or a specific body site or region; and

[0881] (e) Optionally compare the amount of the compound / protein aggregate complex with the normal control value.

[0882] The steps (a) through (e) have been explained above.

[0883] In the method for predicting responsiveness, the method may further include steps (i) to (vi) before step (a):

[0884] (i) Exposing a sample or a specific body part or area suspected of containing α-synuclein aggregates (including but not limited to Lewy bodies and / or Lewy neurites) to the compound of the present invention or a compound of the present invention or a detectable labeled thereof, stereoisomer, racemic mixture, pharmaceutically acceptable salt, hydrate or solvate, said compound specifically binding to α-synuclein aggregates (including but not limited to Lewy bodies and / or Lewy neurites);

[0885] (ii) To bind the compound to α-synuclein aggregates (including but not limited to Lewy bodies and / or Lewy neurites) to form a compound / (α-synuclein aggregates (including but not limited to Lewy bodies and / or Lewy neurites)) complex;

[0886] (iii) Detect the formation of the compound / α-synuclein aggregate (including but not limited to Lewy bodies and / or Lewy neurites) complex;

[0887] (iv) Establish a correlation between the presence or absence of the compound / α-synuclein aggregate (including but not limited to Lewy bodies and / or Lewy neurites) complex and the presence or absence of α-synuclein aggregate (including but not limited to Lewy bodies and / or Lewy neurites) in the sample or a specific body part or region.

[0888] (v) Optionally, compare the amount of the compound / / (α-synuclein aggregates (including but not limited to Lewy bodies and / or Lewy neurites)) with normal control values; and

[0889] (vi) Treat patients with medication.

[0890] Optionally, the method may further include step (A) after step (d) or step (e):

[0891] (A) Compare the amount of compound / (α-synuclein aggregate (including but not limited to Lewy body and / or Lewy neurite)) complex determined in step (iv) with the amount of compound / (α-synuclein aggregate (including but not limited to Lewy body and / or Lewy neurite)) complex determined in step (d).

[0892] To determine the responsiveness over time, steps (a) to (c) and optional steps (d) and (e) of the method for predicting responsiveness may be repeated once or multiple times.

[0893] In methods for predicting responsiveness, the amount of the compound / protein aggregate complex can optionally be compared at various time points during treatment, such as before and after treatment begins, or at various time points after treatment begins. Changes in the amount of the compound / protein aggregate complex, particularly a decrease, can indicate a high potential for response to the corresponding treatment.

[0894] Optionally, diagnostic compositions may be used before, during, and after surgical procedures (e.g., deep brain stimulation (DBS)) and noninvasive brain stimulation (e.g., repetitive transcranial magnetic stimulation (rTMS)) to visualize α-synuclein aggregates before, during, and after such procedures. Surgical techniques (including DBS) improve late-stage symptoms of PD on top of currently best-used medical therapies. Over the past 20 years, RTMS has been closely examined as a potential treatment for PD (Ying-hui Chou et al. JAMA Neurol. 2015 Apr 1; ​​72(4): 432-440).

[0895] In another embodiment of the invention, the diagnostic composition can be used in a method for collecting data on residual disease, condition, or abnormality in a patient suffering from a disease, condition, or abnormality associated with α-synuclein aggregates (including but not limited to Lewy bodies and / or Lewy neurites), the patient having been treated with surgical procedures or non-invasive brain stimulation procedures, wherein the method comprises the following steps:

[0896] (a) Exposing a sample or a specific body part or region suspected of containing α-synuclein aggregates (including but not limited to Lewy bodies and / or Lewy neurites) to the compound of the present invention or a detectable labeled compound thereof, stereoisomer, racemic mixture, pharmaceutically acceptable salt, hydrate or solvate, said compound specifically binding to α-synuclein aggregates (including but not limited to Lewy bodies and / or Lewy neurites);

[0897] (b) To bind the compound to α-synuclein aggregates (including but not limited to Lewy bodies and / or Lewy neurites) to form a compound / (α-synuclein aggregates (including but not limited to Lewy bodies and / or Lewy neurites)) complex;

[0898] (c) Detect the formation of the compound / (α-synuclein aggregates (including but not limited to Lewy bodies and / or Lewy neurites)) complex;

[0899] (d) Optionally, establish a correlation between the presence or absence of the compound / (α-synuclein aggregate (including but not limited to Lewy bodies and / or Lewy neurites)) complex and the presence or absence of α-synuclein aggregates (including but not limited to Lewy bodies and / or Lewy neurites) in the sample or a specific body part or region; and

[0900] (e) Optionally compare the amount of the compound / (α-synuclein aggregate (including but not limited to Lewy bodies and / or Lewy neurites)) complex with normal control values.

[0901] It should be understood that the term “monitoring minimal residual disease” as used herein refers to monitoring the evolution of a disease. For example, monitoring the evolution of a disease, condition, or abnormality in patients with a disease, condition, or abnormality associated with α-synuclein aggregates (including but not limited to Lewy bodies and / or Lewy neurites).

[0902] The compounds of the present invention, or their detectable labeled compounds, stereoisomers, racemic mixtures, pharmaceutically acceptable salts, hydrates, or solvates, or their precursors, may also be incorporated into test kits for detecting α-synuclein protein aggregates (including, but not limited to, Lewy bodies and / or Lewy neurites). The test kit typically comprises a container holding one or more of the compounds of the present invention, or their detectable labeled compounds, stereoisomers, racemic mixtures, pharmaceutically acceptable salts, hydrates, or solvates, or their precursors, and instructions for using said compound to bind α-synuclein aggregates (including, but not limited to, Lewy bodies and / or Lewy neurites) to form a compound / protein aggregate complex and for detecting the formation of said compound / protein aggregate complex, such that the presence or absence of said compound / protein aggregate complex is correlated with the presence or absence of said α-synuclein aggregates (including, but not limited to, Lewy bodies and / or Lewy neurites).

[0903] The term “test kit” generally refers to any diagnostic kit known in the art. More specifically, the latter term refers to diagnostic kits such as those described by Zrein et al. in Clin. Diagn. Lab. Immunol., 1998, 5, 45-49.

[0904] The identifiable labeled compounds of the present invention or their stereoisomers, racemic mixtures, pharmaceutically acceptable salts, hydrates or solvates, preferably using 18 The dosage of the F-labeled compound of formula (III-F) will vary depending on the exact compound to be administered, the patient's weight, the size and type of the sample, and other variables that are obvious to those skilled in the art. Typically, the dosage is preferably in the range of 0.001 μg / kg to 10 μg / kg, more preferably 0.01 μg / kg to 1.0 μg / kg. The radioactive dose can be, for example, 100-600 MBq, more preferably 150-450 MBq.

[0905] In another embodiment, the present invention provides a method for imaging diseases, conditions, or abnormalities associated with α-synuclein aggregates (including but not limited to Lewy bodies and / or Lewy neurites) in a sample or a specific body part or region, particularly in the brain or a sample taken from a patient's brain, the method comprising the following steps:

[0906] (a) Contact a sample or specific body part or region suspected of containing α-synuclein aggregates (including but not limited to Lewy bodies and / or Lewy neurites) with a compound of formula (I) or its subforms (e.g. (IIa), (IIb), (IIIa), (IIIb), (IIIc), (III-F), (III-H)) or its detectable labeled compounds, stereoisomers, racemic mixtures, pharmaceutically acceptable salts, hydrates or solvates;

[0907] (b) to bind the compound to α-synuclein aggregates (including but not limited to Lewy bodies and / or Lewy neurites); and

[0908] (c) Image the sample, a specific body part or body region using an imaging system.

[0909] In another embodiment, the present invention provides a method for determining the amount of α-synuclein aggregates (including but not limited to Lewy bodies and / or Lewy neurites) in a sample or a specific body part or region, the method comprising the following steps:

[0910] (a) Contact a sample or specific body part or region suspected of containing α-synuclein aggregates (including but not limited to Lewy bodies and / or Lewy neurites) with a compound of formula (I) or its subforms (e.g. (IIa), (IIb), (IIIa), (IIIb), (IIIc), (III-F), (III-H)) or its detectable labeled compounds, stereoisomers, racemic mixtures, pharmaceutically acceptable salts, hydrates or solvates;

[0911] (b) To bind the compound to α-synuclein aggregates (including but not limited to Lewy bodies and / or Lewy neurites);

[0912] (c) Detecting compounds that bind to α-synuclein aggregates (including but not limited to Lewy bodies and / or Lewy neurites);

[0913] (d) Determine the amount of compounds that bind to α-synuclein aggregates (including but not limited to Lewy bodies and / or Lewy neurites); and

[0914] (e) Optionally calculate the amount of α-synuclein aggregates (including, but not limited to, Lewy bodies and / or Lewy neurites) in a sample, a specific body part, or a body region.

[0915] In another embodiment, the present invention provides a method for diagnosing diseases, conditions, or abnormalities associated with α-synuclein aggregates (including but not limited to Lewy bodies and / or Lewy neurites), the method comprising the following steps:

[0916] (a) Contact a sample or specific body part or region suspected of containing α-synuclein aggregates (including but not limited to Lewy bodies and / or Lewy neurites) with a compound of formula (I) or its subforms (e.g. (IIa), (IIb), (IIIa), (IIIb), (IIIc), (III-F), (III-H)) or its detectable labeled compounds, stereoisomers, racemic mixtures, pharmaceutically acceptable salts, hydrates or solvates;

[0917] (b) To bind the compound to α-synuclein aggregates (including but not limited to Lewy bodies and / or Lewy neurites);

[0918] (c) Detection of compounds that bind to α-synuclein aggregates (including but not limited to Lewy bodies and / or Lewy neurites); and

[0919] (d) Establish a correlation between the presence or absence of compounds that bind to α-synuclein aggregates (including but not limited to Lewy bodies and / or Lewy neurites) and diseases, conditions or abnormalities associated with α-synuclein aggregates (including but not limited to Lewy bodies and / or Lewy neurites).

[0920] In another embodiment, the present invention provides a method for collecting data for diagnosing diseases, conditions, or abnormalities associated with α-synuclein aggregates (including but not limited to Lewy bodies and / or Lewy neurites), the method comprising the following steps:

[0921] (a) Contact a sample or specific body part or region suspected of containing α-synuclein aggregates (including but not limited to Lewy bodies and / or Lewy neurites) with a compound of formula (I) or its subforms (e.g. (IIa), (IIb), (IIIa), (IIIb), (IIIc), (III-F), (III-H)) or its detectable labeled compounds, stereoisomers, racemic mixtures, pharmaceutically acceptable salts, hydrates or solvates;

[0922] (b) To bind the compound to α-synuclein aggregates (including but not limited to Lewy bodies and / or Lewy neurites);

[0923] (c) Detecting compounds that bind to α-synuclein aggregates (including but not limited to Lewy bodies and / or Lewy neurites); and

[0924] (d) Optionally, a correlation is established between the presence or absence of a compound that binds to α-synuclein aggregates (including but not limited to Lewy bodies and / or Lewy neurites) and the presence or absence of α-synuclein aggregates (including but not limited to Lewy bodies and / or Lewy neurites) in a sample or a specific body part or region.

[0925] In another embodiment, the present invention provides a method for collecting data on susceptibility to diseases, conditions, or abnormalities associated with α-synuclein aggregates (including but not limited to Lewy bodies and / or Lewy neurites), the method comprising the following steps:

[0926] (a) Contact a sample or specific body part or region suspected of containing α-synuclein aggregates (including but not limited to Lewy bodies and / or Lewy neurites) with a compound of formula (I) or its subforms (e.g. (IIa), (IIb), (IIIa), (IIIb), (IIIc), (III-F), (III-H)) or its detectable labeled compounds, stereoisomers, racemic mixtures, pharmaceutically acceptable salts, hydrates or solvates;

[0927] (b) To bind the compound to α-synuclein aggregates (including but not limited to Lewy bodies and / or Lewy neurites);

[0928] (c) Detecting compounds that bind to α-synuclein aggregates (including but not limited to Lewy bodies and / or Lewy neurites); and

[0929] (d) Optionally, a correlation is established between the presence or absence of a compound that binds to α-synuclein aggregates (including but not limited to Lewy bodies and / or Lewy neurites) and the presence or absence of α-synuclein aggregates (including but not limited to Lewy bodies and / or Lewy neurites) in a sample or a specific body part or region.

[0930] If the amount of compounds binding to α-synuclein aggregates is higher than the normal control value in healthy / reference individuals, this indicates that the patient has a disease, condition, or abnormality associated with α-synuclein aggregates or is at risk of developing such a disease, condition, or abnormality. Specifically, if the amount of compounds binding to α-synuclein aggregates is higher than the amount expected in individuals without clinical evidence of neurodegenerative disease, it can be presumed that the patient has a susceptibility to a disease, condition, or abnormality associated with α-synuclein aggregates or synucleinopathy.

[0931] In another embodiment, the present invention provides a method for collecting data for the prognosis of diseases, conditions, or abnormalities associated with α-synuclein aggregates (including but not limited to Lewy bodies and / or Lewy neurites), wherein the method comprises the following steps:

[0932] (a) To contact a sample, specific body part or region suspected of containing α-synuclein aggregates (including but not limited to Lewy bodies and / or Lewy neurites) with a compound of formula (I) or its subforms (e.g. (IIa), (IIb), (IIIa), (IIIb), (IIIc), (III-F), (III-H)) or its detectable labeled compounds, stereoisomers, racemic mixtures, pharmaceutically acceptable salts, hydrates or solvates;

[0933] (b) To bind the compound to α-synuclein aggregates (including but not limited to Lewy bodies and / or Lewy neurites);

[0934] (c) Detecting compounds that bind to α-synuclein aggregates (including but not limited to Lewy bodies and / or Lewy neurites);

[0935] (d) Optionally, establish a correlation between the presence or absence of compounds that bind to α-synuclein aggregates (including but not limited to Lewy bodies and / or Lewy neurites) and the presence or absence of α-synuclein aggregates (including but not limited to Lewy bodies and / or Lewy neurites) in the sample or a specific body part or region; and

[0936] (e) Optionally repeat steps (a) through (c) and optional step (d) (if any) at least once.

[0937] The prospect of progression and / or recovery of the disease, condition, or abnormality (e.g., probability, duration, and / or extent) can be estimated by a medical practitioner based on the presence or absence of a compound that binds to α-synuclein aggregates, the amount of the compound that binds to α-synuclein aggregates, etc. If desired, steps (a) through (c) and optional step (d) (if present) can be repeated over time to monitor the progression of the disease, condition, or abnormality, thereby allowing for more reliable estimation.

[0938] In another embodiment, the present invention provides a method for collecting data for monitoring disease progression in patients with diseases, conditions, or abnormalities associated with α-synuclein aggregates (including but not limited to Lewy bodies and / or Lewy neurites), the method comprising the following steps:

[0939] (a) To contact a sample, specific body part or region suspected of containing α-synuclein aggregates (including but not limited to Lewy bodies and / or Lewy neurites) with a compound of formula (I) or its subforms (e.g. (IIa), (IIb), (IIIa), (IIIb), (IIIc), (III-F), (III-H)) or its detectable labeled compounds, stereoisomers, racemic mixtures, pharmaceutically acceptable salts, hydrates or solvates;

[0940] (b) To bind the compound to α-synuclein aggregates (including but not limited to Lewy bodies and / or Lewy neurites);

[0941] (c) Detecting compounds that bind to α-synuclein aggregates (including but not limited to Lewy bodies and / or Lewy neurites);

[0942] (d) Optionally, establish a correlation between the presence or absence of compounds that bind to α-synuclein aggregates (including but not limited to Lewy bodies and / or Lewy neurites) and the presence or absence of α-synuclein aggregates (including but not limited to Lewy bodies and / or Lewy neurites) in the sample or a specific body part or region; and

[0943] (e) Optionally repeat steps (a) through (c) and optional step (d) (if any) at least once.

[0944] Typically, the patient is currently undergoing or has been undergoing treatment for a disease, condition, or abnormality associated with α-synuclein aggregates, or is currently / has undergone treatment for synucleinopathy. In particular, treatment may involve the administration of a medication suitable for treating a disease, condition, or abnormality associated with α-synuclein aggregates.

[0945] In another embodiment, the present invention provides a method for collecting data for monitoring the progression of a disease, condition, or abnormality associated with α-synuclein aggregates (including but not limited to Lewy bodies and / or Lewy neurites) in a patient, the method comprising the following steps:

[0946] (a) To contact a sample, specific body part or region suspected of containing α-synuclein aggregates (including but not limited to Lewy bodies and / or Lewy neurites) with a compound of formula (I) or its subforms (e.g. (IIa), (IIb), (IIIa), (IIIb), (IIIc), (III-F), (III-H)) or its detectable labeled compounds, stereoisomers, racemic mixtures, pharmaceutically acceptable salts, hydrates or solvates;

[0947] (b) To bind the compound to α-synuclein aggregates (including but not limited to Lewy bodies and / or Lewy neurites);

[0948] (c) Detecting compounds that bind to α-synuclein aggregates (including but not limited to Lewy bodies and / or Lewy neurites);

[0949] (d) Optionally, establish a correlation between the presence or absence of compounds that bind to α-synuclein aggregates (including but not limited to Lewy bodies and / or Lewy neurites) and the presence or absence of α-synuclein aggregates (including but not limited to Lewy bodies and / or Lewy neurites) in the sample or a specific body part or region; and

[0950] (e) Optionally repeat steps (a) through (c) and optional step (d) (if any) at least once.

[0951] Typically, the patient is currently undergoing or has been undergoing treatment for a disease, condition, or abnormality associated with α-synuclein aggregates, or is currently undergoing or has been undergoing treatment for synucleinopathy. In particular, treatment may involve the administration of a medication suitable for treating a disease, condition, or abnormality associated with α-synuclein aggregates.

[0952] In another embodiment, the present invention provides a method for acquiring data to predict the responsiveness of a patient suffering from a disease, condition, or abnormality associated with α-synuclein aggregates (including but not limited to Lewy bodies and / or Lewy neurites) to treatments associated with α-synuclein aggregates (including but not limited to Lewy bodies and / or Lewy neurites), the method comprising the following steps:

[0953] (a) To contact a sample, specific body part or region suspected of containing α-synuclein aggregates (including but not limited to Lewy bodies and / or Lewy neurites) with a compound of formula (I) or its subforms (e.g. (IIa), (IIb), (IIIa), (IIIb), (IIIc), (III-F), (III-H)) or its detectable labeled compounds, stereoisomers, racemic mixtures, pharmaceutically acceptable salts, hydrates or solvates;

[0954] (b) To bind the compound to α-synuclein aggregates (including but not limited to Lewy bodies and / or Lewy neurites);

[0955] (c) Detecting compounds that bind to α-synuclein aggregates (including but not limited to Lewy bodies and / or Lewy neurites);

[0956] (d) Optionally, establish a correlation between the presence or absence of compounds that bind to α-synuclein aggregates (including but not limited to Lewy bodies and / or Lewy neurites) and the presence or absence of α-synuclein aggregates (including but not limited to Lewy bodies and / or Lewy neurites) in the sample or a specific body part or region; and

[0957] (e) Optionally repeat steps (a) through (c) and optional step (d) (if any) at least once.

[0958] Typically, the patient is currently undergoing or has been undergoing treatment for a disease, condition, or abnormality associated with α-synuclein aggregates, or is currently undergoing or has been undergoing treatment for synucleinopathy. In particular, treatment may involve the administration of medications suitable for treating diseases, conditions, or abnormalities associated with α-synuclein aggregates.

[0959] If the amount of compound binding to α-synuclein aggregates decreases over time, it can be presumed that the patient is responding to treatment. If the amount of compound binding to α-synuclein aggregates remains substantially constant or increases over time, it can be presumed that the patient is not responding to treatment.

[0960] Alternatively, responsiveness can be estimated by determining the amount of compound that binds to α-synuclein aggregates. The amount of compound binding to α-synuclein aggregates can be compared to control values, such as normal controls, preclinical controls, or clinical controls. Alternatively, control values ​​can refer to those of individuals known to respond to a treatment, or those of individuals known to be unresponsive to a treatment. Results regarding responsiveness can be "responsive" to a therapy, "unresponsive" to a therapy, or "response indeterminate." Response to treatment may vary from patient to patient.

[0961] In another embodiment, the present invention provides a method as defined herein, wherein the step of optionally establishing a correlation between the presence or absence of a compound that binds to α-synuclein aggregates (including, but not limited to, Lewy bodies and / or Lewy neurites) and the presence or absence of α-synuclein aggregates (including, but not limited to, Lewy bodies and / or Lewy neurites) in a sample or a specific body part or region comprises:

[0962] - Determine the amount of compounds that bind to α-synuclein aggregates (including but not limited to Lewy bodies and / or Lewy neurites);

[0963] - Establish a correlation between the amount of compounds binding to α-synuclein aggregates (including but not limited to Lewy bodies and / or Lewy neurites) and the amount of α-synuclein aggregates (including but not limited to Lewy bodies and / or Lewy neurites) in a sample or a specific body part or region; and

[0964] -Optionally compare the amount of α-synuclein aggregates (including but not limited to Lewy bodies and / or Lewy neurites) in a sample or a specific body part or region with normal control values ​​in healthy control individuals.

[0965] Control values ​​can be, for example, normal control values, preclinical control values, and / or clinical control values.

[0966] “Healthy control individuals” or “healthy volunteer (HV) individuals” are people who do not show clinical evidence of neurodegenerative disease. These are selected as defined in Section 15, “First-in-Human (FIH) Studies,” of the “Bioassay Description and Corresponding Results” section of this paper.

[0967] If, in any of the methods summarized above, the amount of compounds binding to α-synuclein aggregates is higher than the normal control value, then the patient may be expected to have or be likely to have a disease, condition, or abnormality associated with α-synuclein aggregates or synucleinopathy.

[0968] Any compound of the present invention can be used in the methods described above. Preferably, detectable labeled compounds of the present invention, as disclosed herein, are used in the methods described above.

[0969] The specific body part or body area is preferably of mammals, more preferably of humans, including whole or part of the body area or body part of a patient suspected of containing α-synuclein aggregates.

[0970] The sample may be selected from tissues or body fluids suspected of containing α-synuclein aggregates, and the sample is obtained from a patient. Preferably, the tissue is selected from brain tissue. Examples of body fluids include cerebrospinal fluid (CSF) or blood. The sample may be obtained from mammals, more preferably humans. Preferably, the sample is an in vitro sample from a patient.

[0971] In in vivo methods, by administering an effective amount of the compound of the present invention to a patient, a specific body site or area can be brought into contact with the compound. An effective amount of the compound of the present invention is a suitable amount that allows the determination, using selected analytical techniques, of the presence or absence of α-synuclein aggregates (including, but not limited to, Lewy bodies and / or Lewy neurites) in a specific body site or area.

[0972] The step of binding the compound to α-synuclein aggregates (including, but not limited to, Lewy bodies and / or Lewy neurites) includes allowing sufficient time for the compound of the present invention to bind to the α-synuclein aggregates (including, but not limited to, Lewy bodies and / or Lewy neurites). The amount of time required for binding will depend on the type of test (e.g., in vitro or in vivo) and can be determined by those skilled in the art through routine experiments. In in vivo methods, the amount of time will depend on the time required for the compound to reach a specific body site or region suspected of containing α-synuclein aggregates (including, but not limited to, Lewy bodies and / or Lewy neurites). The amount of time should not be excessive to avoid washout and / or metabolism of the compound of the present invention.

[0973] There are no particular limitations on the methods for detecting compounds that bind to α-synuclein aggregates (including but not limited to Lewy bodies and / or Lewy neurites), but rather depend on the detectable markers, sample type, specific body site or region, and whether the method is in vitro or in vivo. Possible detection methods include, but are not limited to, fluorescence imaging techniques or nuclear imaging techniques, such as positron emission tomography (PET), single-photon emission computed tomography (SPECT), magnetic resonance imaging (MRI), and contrast-enhanced magnetic resonance imaging (MRI). Fluorescence imaging techniques and / or nuclear imaging techniques can be used to monitor and / or visualize the distribution of the compounds of the present invention in a sample or in vivo. The imaging system thus provides images of the bound detectable markers (e.g., radioactive isotopes, particularly positron emitters or gamma emitters) present in the test sample, the specific body site being tested, or the body region being tested. Preferably, compounds that bind to α-synuclein aggregates (including but not limited to Lewy bodies and / or Lewy neurites) are detected by imaging devices such as PET or SPECT scanners.

[0974] The amount of compounds that bind to α-synuclein aggregates (including but not limited to Lewy bodies and / or Lewy neurites) can be determined by visual or quantitative analysis, such as using PET scan images.

[0975] In any of the methods described above, steps (a) through (c) and optional step (d) (if present) may be repeated at least once. Repetition of steps is particularly useful in methods for collecting data for prognosis, for collecting data for monitoring disease progression, for collecting data for monitoring progression, and for collecting data for predicting responsiveness. In these methods, it may be advantageous to monitor the patient over time and repeat the above steps after a certain period of time. The time interval before repeating the above steps can be determined by the clinician based on the severity of the disease, condition, or abnormality associated with α-synuclein aggregates (including but not limited to Lewy bodies and / or Lewy neurites) or synucleinopathy.

[0976] In another aspect, the present invention relates to a method for imaging diseases, conditions, or abnormalities associated with α-synuclein aggregates (including but not limited to Lewy bodies and / or Lewy neurites) in an individual, the method comprising the following steps:

[0977] (a) administering to an individual a compound of formula (I) or its subforms (e.g. (IIa), (IIb), (IIIa), (IIIb), (IIIc), (III-F), (III-H)) or a detectable labeled compound thereof, stereoisomer, racemic mixture, pharmaceutically acceptable salt, hydrate or solvate;

[0978] (b) to bind the compound to α-synuclein aggregates (including but not limited to Lewy bodies and / or Lewy neurites); and

[0979] (c) Detecting compounds that bind to α-synuclein aggregates, including but not limited to Lewy bodies and / or Lewy neurites.

[0980] In another aspect, the present invention relates to a method for imaging diseases, conditions, or abnormalities associated with α-synuclein aggregates (including but not limited to Lewy bodies and / or Lewy neurites) in an individual, the method comprising the following steps:

[0981] (a) administering to an individual a compound of formula (I) or its subforms (e.g., (IIa), (IIb), (IIIa), (IIIb), (IIIc), (III-F), (III-H)) or a detectably labeled compound thereof, stereoisomer, racemic mixture, pharmaceutically acceptable salt, hydrate, or solvate; and

[0982] (b) To enable brain imaging of an individual.

[0983] When the compound binds to α-synuclein aggregates (including but not limited to Lewy bodies and / or Lewy neurites), the individual's brain should be imaged. The compound binding to α-synuclein aggregates (including but not limited to Lewy bodies and / or Lewy neurites) should then be imaged in the individual's brain.

[0984] In another aspect, the present invention relates to a method for positron emission tomography (PET) imaging of α-synuclein aggregates (including but not limited to Lewy bodies and / or Lewy neurites) in individual tissues, the method comprising the following steps:

[0985] (a) administering to an individual a compound of formula (I) or its subforms (e.g. (IIa), (IIb), (IIIa), (IIIb), (IIIc), (III-F), (III-H)) or a detectable labeled compound thereof, stereoisomer, racemic mixture, pharmaceutically acceptable salt, hydrate or solvate;

[0986] (b) to allow the compound to penetrate into the tissues of an individual; and

[0987] (c) Acquiring positron emission tomography (PET) images of individual tissues;

[0988] The tissue is central nervous system (CNS) tissue, eye or brain tissue, preferably brain tissue.

[0989] PET imaging should be performed when the compound penetrates into the tissue and binds to α-synuclein aggregates (including but not limited to Lewy bodies and / or Lewy neurites).

[0990] In another aspect, the present invention relates to a method for detecting neurological diseases, conditions, or abnormalities associated with an individual's α-synuclein aggregates (including but not limited to Lewy bodies and / or Lewy neurites), the method comprising the following steps:

[0991] (a) administering to an individual a compound of formula (I) or its subforms (e.g. (IIa), (IIb), (IIIa), (IIIb), (IIIc), (III-F), (III-H)) or a detectable labeled compound thereof, stereoisomer, racemic mixture, pharmaceutically acceptable salt, hydrate or solvate;

[0992] (b) to bind the compound to α-synuclein aggregates (including but not limited to Lewy bodies and / or Lewy neurites); and

[0993] (c) Determine the radioactive signal of the compound, which binds to α-synuclein aggregates (including but not limited to Lewy bodies and / or Lewy neurites).

[0994] When containing at least one radiolabeled atom (e.g.) 3 H, 2 H or 18 F) The present invention can detect the observation of radioactive signals as mentioned herein when labeled compounds bind to α-synuclein aggregates (including but not limited to Lewy bodies and / or Lewy neurites).

[0995] In another aspect, the present invention relates to a method (e.g., in vivo or in vitro method) for detecting and / or quantifying α-synuclein aggregates (including but not limited to Lewy bodies and / or Lewy neurites) in individual tissues, the method comprising the following steps:

[0996] (a) To contact the tissue of an individual with a compound of formula (I) or its subforms (e.g. (IIa), (IIb), (IIIa), (IIIb), (IIIc), (III-F), (III-H)) or a detectable labeled compound thereof, stereoisomer, racemic mixture, pharmaceutically acceptable salt, hydrate or solvate;

[0997] (b) to bind the compound to α-synuclein aggregates (including but not limited to Lewy bodies and / or Lewy neurites); and

[0998] (c) Detection and / or quantification of compounds that bind to α-synuclein aggregates (including but not limited to Lewy bodies and / or Lewy neurites) using positron emission tomography.

[0999] In another aspect, the present invention relates to a method for diagnostic imaging of an individual's brain, the method comprising the following steps:

[1000] (a) administering to an individual a compound of formula (I) or its subforms (e.g., (IIa), (IIb), (IIIa), (IIIb), (IIIc), (III-F), (III-H)) or a detectably labeled compound thereof, stereoisomer, racemic mixture, pharmaceutically acceptable salt, hydrate, or solvate; and

[1001] (b) Obtain brain images of an individual using positron emission tomography.

[1002] In the method of the present invention, compounds of formula (I) or its subforms (e.g., (IIa), (IIb), (IIIa), (IIIb), (IIIc), (III-F), (III-H)) or compounds with detectable labels thereof, stereoisomers, racemic mixtures, pharmaceutically acceptable salts, hydrates, or solvates are typically applied in a detectable amount, i.e., an amount detectable by the apparatus that can be used to detect the compound in the corresponding method. There is no particular limitation on this amount, but it depends on the compound of formula (I), the type of detectable label, the sensitivity of the corresponding analytical method, and the corresponding apparatus. This amount can be appropriately selected by those skilled in the art.

[1003] Radiopharmaceutical preparations

[1004] The compounds of the present invention, or their detectable labeled compounds, stereoisomers, racemic mixtures, pharmaceutically acceptable salts, hydrates, or solvates, preferably of formula (I) or its subforms (e.g., (IIa), (IIb), (IIIa), (IIIb), (IIIc), (III-F), (III-H)), can also be used in kits for preparing radiopharmaceutical formulations. Radiopharmaceuticals are typically prepared immediately before use due to radioactive decay. The kit typically comprises a precursor of the compound of the present invention, or its detectable labeled compounds, stereoisomers, racemic mixtures, pharmaceutically acceptable salts, hydrates, or solvates, and a reagent for reacting with said precursor to introduce a radiolabel into the compound of the present invention, or its detectable labeled compounds, stereoisomers, racemic mixtures, pharmaceutically acceptable salts, hydrates, or solvates. The precursors of the compound of the present invention, or their detectable labeled compounds, stereoisomers, racemic mixtures, pharmaceutically acceptable salts, hydrates, or solvates, can be, for example, compounds having formulas (IV-F), (IV-H), or (IV-J). The reagent can be a radioactive label, for example... 18 F or 3 H reagent.

[1005] Pharmaceutical Composition

[1006] The compounds of the present invention or their detectable and labelable compounds, stereoisomers, racemic mixtures, pharmaceutically acceptable salts, hydrates or solvates may be used to treat, prevent or alleviate diseases, symptoms or abnormalities associated with α-synuclein aggregates.

[1007] The compounds of the present invention, or detectable and labelable compounds thereof, stereoisomers, racemic mixtures, pharmaceutically acceptable salts, hydrates, or solvates, preferably compounds of formula (I), are suitable for the treatment, prevention, or relief of diseases, symptoms, or abnormalities associated with α-synuclein aggregates (including, but not limited to, Lewy bodies and / or Lewy neurites). Diseases involving α-synuclein aggregates are generally classified as synucleinopathy (or α-synucleinopathy). The compounds of this invention, or detectable and labelable compounds thereof, stereoisomers, racemic mixtures, pharmaceutically acceptable salts, hydrates, or solvates, are suitable for the treatment, prevention, or alleviation of diseases, symptoms, or abnormalities, including but not limited to Parkinson's disease (sporadic, familial with α-synuclein mutations or familial with mutations other than α-synuclein, simple autonomic failure, and Lewy body dysphagia), SNCA repeat carriers, Lewy body dementia (“simple” Lewy body dementia), Alzheimer's disease, sporadic Alzheimer's disease, familial Alzheimer's disease with APP mutations, familial Alzheimer's disease with PS-1, PS-2, or other mutations, familial British dementia, Lewy body variants of Alzheimer's disease, and normal aging in Down syndrome. Synucleinic diseases involving α-synuclein aggregates of neurons and glial cells include, but are not limited to, multiple system atrophy (MSA) (Shy-Drager syndrome, striatal substantia nigra, and olivary pontocerebellar atrophy). Other diseases that may involve α-synuclein immune response disorders include traumatic brain injury, chronic traumatic encephalopathy, tau disorders (Pick disease, frontotemporal dementia, progressive supranuclear palsy, corticobasal degeneration, and Niemann-Pick type C1 disease), motor neuron disease, amyotrophic lateral sclerosis (sporadic, familial, and Guam ALS-dementia syndrome), axillary dystrophy, neurodegeneration with type 1 brain iron accumulation (Has-Schwarz syndrome), prions, ataxia-telangiectasia, idiopathic orofacial motor disorder, subacute sclerosing panencephalitis, Gaucher disease, and other lysosomal storage disorders (including Kufor-Rakeb syndrome and Sanfilippo syndrome) and tachyoamyotrophic lateral sclerosis (REM) sleep behavior disorders.(Jellinger, Mov Disord 2003, 18 Supplement 6, S2-12; Galvin et al. JAMA Neurology 2001, 58(2), 186-190; Kovari et al. Acta Neuropathol. 2007, 114(3), 295-8; Saito et al. J Neuropathol Exp Neurol. 2004, 63(4), 323-328; McKee et al. Brain, 2013, 136(Pt 1), 43-64; Puschmann et al. Parkinsonism Relat Disord 2012, 18S1, S24-S27; Usenovic et al. J Neurosci. 2012, 32(12), 4240-4246; Winder-Rhodes et al. Mov Disord. 2012, 27(2), 312-315; Ferman et al., J Int Neuropsychol Soc. 2002, 8(7), 907-914). Preferably, the compounds of the present invention are suitable for treating, preventing or alleviating Parkinson's disease (PD).

[1008] In pharmaceutical applications, the compounds of the present invention, or their detectable-labeled compounds, stereoisomers, racemic mixtures, pharmaceutically acceptable salts, hydrates, or solvates thereof, are preferably administered in the form of pharmaceutical compositions comprising the compounds of the present invention. In this invention, a "pharmaceutical composition" is defined as a composition comprising one or more compounds of the present invention, or their detectable-labeled compounds, stereoisomers, racemic mixtures, pharmaceutically acceptable salts, hydrates, or solvates thereof, in a form suitable for administration to a patient, such as a mammal, such as a human, and suitable for treating, alleviating, or preventing the specific disease, symptom, or abnormality described herein. Preferably, the pharmaceutical composition further comprises a physiologically acceptable carrier, diluent, adjuvant, or excipient. The dosage of the compounds of the present invention, or their detectable-labeled compounds, stereoisomers, racemic mixtures, pharmaceutically acceptable salts, hydrates, or solvates thereof, can vary depending on the specific compound administered, the patient's weight, and other variables readily apparent to a clinician skilled in the art.

[1009] Although the compounds of the present invention or their detectable-labeled compounds, stereoisomers, racemic mixtures, pharmaceutically acceptable salts, hydrates, or solvates may be administered alone, they are preferably formulated into pharmaceutical compositions according to standard pharmaceutical practice. Therefore, the present invention also provides pharmaceutical compositions comprising a therapeutically effective amount of a compound of formula (I) or its detectable-labeled compounds, stereoisomers, racemic mixtures, pharmaceutically acceptable salts, hydrates, or solvates, and optionally at least one pharmaceutically acceptable excipient, carrier, diluent, or adjuvant.

[1010] Pharmaceutically acceptable excipients are well-known in the pharmaceutical field and, for example, described in Remington's Pharmaceutical Sciences, 15th edition, Mack Publishing Co., New Jersey (1975). Pharmaceutical excipients can be selected based on the intended route of administration and standard pharmaceutical practice. Excipients must be acceptable in the sense that they are harmless to the recipient.

[1011] Pharmaceutically useful excipients that can be used to formulate the pharmaceutical compositions of the present invention or their detectable labeled compounds, stereoisomers, racemic mixtures, pharmaceutically acceptable salts, hydrates, or solvates may include, for example, carriers, solvents, diluents, solvents such as monohydric alcohols such as ethanol and isopropanol, and polyhydric alcohols such as glycols, and edible oils such as soybean oil, coconut oil, olive oil, safflower oil, and cottonseed oil, oleic esters such as ethyl oleate and isopropyl myristate, binders, adjuvants, solubilizers, etc. Thickeners, stabilizers, disintegrants, flow aids, lubricants, buffers, emulsifiers, wetting agents, suspending agents, sweeteners, colorants, flavorings, coatings, preservatives, antioxidants, processing aids, drug delivery modifiers and enhancers, such as calcium phosphate, magnesium stearate, talc, monosaccharides, disaccharides, starch, gelatin, cellulose, methylcellulose, sodium carboxymethylcellulose, dextrose, hydroxypropyl-β-cyclodextrin, polyvinylpyrrolidone, low-melting-point waxes and ion exchange resins.

[1012] The compounds of the present invention, or detectable and labeled compounds thereof, stereoisomers, racemic mixtures, pharmaceutically acceptable salts, hydrates or solvates, and their precursors, can be synthesized by one of the general methods shown in the following embodiments. These methods are given for illustrative purposes only and should not be considered as limiting.

[1013] abbreviation meaning DMFDMA N,N-Dimethylformamide dimethylacetal SNAr Nucleophilic aromatic substitution CsF cesium fluoride DMSO Dimethyl sulfoxide NBS N-bromosuccinimide LG Leaving group WFI Water for Injection HPLC High performance liquid chromatography SPE solid phase extraction

[1014] General synthetic protocols for preparing the compounds and precursors of the present invention:

[1015] Option 1

[1016]

[1017] Commercially available hydrazine can be condensed with a suitable ketone to give the corresponding hydrazone. The crude hydrazone is cyclized using DMF / DMA to give intermediate A. SNAr reaction can be performed using a suitable nucleophile in a suitable solvent and base to give intermediate B. Alternatively, thermal conditions can be applied without a metal catalyst. Deprotection under suitable conditions yields intermediate C. Finally, intermediate C can be further functionalized using palladium-catalyzed amidation or a Ullmann reaction to give compounds of formula (I) or its subforms (e.g., (IIa), (IIb), (IIIa), (IIIb), (IIIc), (III-F), (III-H)). In this example, the starting material comprises R... 0 Let H be the value of R. The general scheme described above applies to R. 0 It is a raw material for C1-C4 alkyl groups.

[1018] Option 1A

[1019]

[1020] An alternative method (Scheme 1A) involves deprotecting intermediate A and then performing an SNAr reaction with a suitable nucleophile, preferably in DMSO in the presence of CsF. Intermediates C and D can be further functionalized, preferably using copper(I) (Ullmann reaction) in the presence of a base and solvent, to give formula (IIIa) and intermediate E. Finally, LG can be introduced into intermediate E to give formula (IV-F). In this example, the starting material contains R. 0 Let H be the value of R. The general scheme described above applies to R. 0 It is a raw material for C1-C4 alkyl groups.

[1021] Option 1B

[1022]

[1023] The general method is described in Scheme 1B according to the same preferred conditions as those described in General Scheme 1 or 1A.

[1024] Intermediate A can be obtained by treating it with hydroxypyrrolidine in a suitable solvent under heating. 18 F-precursor. R 4 The group can be introduced via palladium-catalyzed amidation or the Ullmann reaction. Finally, the alcohol intermediate E can be modified with a leaving group under standard conditions to obtain compounds of formula (IV-F).

[1025] 3 The H- precursor can be converted to a suitable R- precursor via palladium-catalyzed amidation or the Ullmann reaction. 4The compound is obtained by introducing a group into intermediate C. Finally, by halogenating pyridine in a suitable solvent, such as NBS, the compound of formula (IV-H) can be obtained.

[1026] 18 General Synthesis of F-labeled Compounds of the Invention

[1027] This can be achieved by reacting the precursor compound described below with... 18 The F-fluorinating agent reaction causes the LG contained in the precursor compound to be... 18 F substitution, thereby preparing 18 F-labeled compounds having formula (I).

[1028] Can be used 18 The reagents, solvents, and conditions for F-fluorination are well known to those skilled in the art (L. Cai, S. Lu, V. Pike, Eur. J. Org. Chem 2008, 2853-2873; J. Fluorine Chem., 27(1985): 177-191; Coenen, Fluorine-18 Labeling Methods: Features and Possibilities of Basic Reactions, (2006): Schubiger PA, Friebe M., Lehmann L., (eds), PET-Chemistry-The Driving Force in Molecular Imaging. Springer, Berlin Heidelberg, pp. 15-50). Preferably, for 18 The solvent for F-fluorination is DMF, DMSO, acetonitrile, DMA or a mixture thereof, preferably acetonitrile or DMSO.

[1029] Any suitable one can be used. 18 F-fluorinating agents. Typical examples include H 18 F, alkali metals or alkaline earth metals 18 F-fluorides (e.g., K) 18 F, Rb 18 F, Cs 18 F and Na 18 F). Optional, 18 F-fluorinating agents can be used in combination with chelating agents, such as cavitation ligands (e.g., 4,7,13,16,21,24-hexaoxa-1,10-diazabicyclo[8.8.8]-hexacosane- ) or crown ether (e.g., 18-crown-6). Or, 18 F-fluorinating agents can be used as... 18 F is a tetraalkylammonium salt or a tetraalkylphosphonium salt; for example18 F's four (C) 1-6 alkyl)ammonium salts or 18 F's four (C) 1-6 Alkyl)phosphonium salt. Preferably, 18 F-fluorinating agent is K 18 F, H 18 F, Cs 18 F, Na 18 F, 18 F's four (C) 1-6 Alkyl ammonium salts, kryptofix

[222] 18 F or tetrabutylammonium fluoride [ 18 F).

[1030] Despite the above regarding 18 F, as a radioactive label, showed a reaction, but other radioactive labels can be introduced in a similar manner.

[1031] The present invention is illustrated by the following examples; however, these examples should not be construed as limiting. Example

[1032] All reagents and solvents were obtained from commercial sources and were ready for use without further purification. Protons were recorded in deuterated solvents on a Bruker DRX-400MHz NMR spectrometer, a Bruker AV-400MHz NMR spectrometer, or a SpinSolve 80MHz NMR spectrometer. 1 H) Spectroscopy. Mass spectra (MS) were recorded on an Advion CMS mass spectrometer or a UPLC H-Class Plus with a photodiode array detector and a QDA mass spectrometer from Waters. Chromatography was performed using silica gel (Fluka: silica gel 60, 0.063–0.2 mm) and suitable solvents as shown in the specific examples. Rapid purification was performed using the Biotage Isolera One rapid purification system, applying an HP-SIL or KP-NH SNAP column (Biotage) and the solvent gradients shown in the specific examples. Thin-layer chromatography (TLC) was performed on silica gel plates with UV detection.

[1033] Preparation Example 1

[1034]

[1035] Step A:

[1036] A suspension of 2-bromo-5-hydrazinopyridine (3.21 g, 17.07 mmol) and tert-butyl 2,4-dioxopyrrolidine-1-carboxylate (3.40 g, 17.07 mmol) in ethanol (150 mL) was refluxed for 3 h and monitored by TLC. The crude product was concentrated under reduced pressure and diluted with dichloromethane and water. The layers were separated, and the aqueous layer was extracted twice with dichloromethane. The combined organic layers were dried over Na₂SO₄, filtered, and concentrated under reduced pressure. The crude product was purified by rapid chromatography (silica, 50 g column, 60-80% ethyl acetate in heptane) to give (E)-4-(2-(6-bromopyridin-3-yl)hydrazide)-2-oxopyrrolidine-1-carboxylate as a brown solid (4.97 g, 79%). 1 H NMR (400MHz, DMSO-d6) δ = 9.22 (s, 1H), 8.41 (s, 1H), 7.89 (d, 1H), 7.40 (d, 1H), 7.11 (dd, 1H), 4.57 (s, 1H), 4.30 (s, 2H), 1.45 (s, 9H). MS: 369.06 [M+H] +

[1037] Step B:

[1038] The compound from step A (3.9 g, 10.56 mmol) was stirred in 1,1-dimethoxy-N,N-dimethylmethylamine (80 mL) at 50 °C for 3 h 15 min. The reaction mixture was concentrated to ~10 mL and ethanol was added. The solid was filtered and washed with a small amount of ethanol to give tert-butyl 2-(6-bromopyridin-3-yl)-4-oxo-4,6-dihydropyrrolo[3,4-c]pyrazole-5(2H)-carboxylate as a light brown powder (2.30 g, 57%). 1 H NMR (400MHz, DMSO-d6) δ = 9.20 (s, 1H), 9.00 (d, 1H), 8.28 (dd, 1H), 7.89 (d, 1H), 4.84 (s, 2H), 1.53 (s, 9H). MS: 324.83 [M-tBu + H] +

[1039] Preparation Examples 1A-1H

[1040] The following preparation examples were prepared using 1,1-dimethoxy-N,N-dimethylmethylamine or N,N-dimethylacetamide dimethylacetal and a suitable hydrazone, as described in Preparation Example 1.

[1041]

[1042]

[1043] Preparation Example A

[1044]

[1045] Preparation Example 1 (1000 mg, 2.64 mmol) was stirred in a 37 mL solution of 4 M HCl in dioxane at room temperature for 1 h 45 min. The solvent was evaporated under reduced pressure, and the solid was dissolved in dichloromethane. A saturated NaHCO3 solution was added, and the aqueous phase was extracted twice with dichloromethane. The combined organic layers were filtered to give 2-(6-bromopyridin-3-yl)-5,6-dihydropyrrolo[3,4-c]pyrazole-4(2H)-one, a light brown solid (682 mg, 93%). 1 H NMR (80MHz, DMSO-d6) δ8.96 (d, 2H), 8.37-8.14 (m, 2H), 7.83 (d, IH), 4.39 (s, 2H). MS: 280.95 [M+H] +

[1046] Each of the embodiments A1-A6

[1047] The following preparation examples were prepared according to the method described in Preparation Example A.

[1048]

[1049]

[1050] Preparation Example 2

[1051]

[1052] In a flask under an argon atmosphere, palladium(II) acetate (41.4 mg, 0.185 mmol) and xantphos (320 mg, 0.554 mmol) were mixed in 1,4-dioxane (18 mL) and heated at 100 °C for a few seconds with a preheating block to form the pd-xantphos complex. (R)-3-fluoropyrrolidine hydrochloride (348 mg, 2.77 mmol), cesium carbonate (1804 mg, 5.54 mmol), and preparation example 1 (700 mg, 1.846 mmol) were added. The flask was degassed, purged with argon three times, and the reaction mixture was heated at 120 °C for 30 min. The reaction mixture was cooled to room temperature, and the residue was dissolved in ethyl acetate and water. The phases were separated, and the aqueous phase was extracted twice. The combined organic layers were dried over Na₂SO₄ and evaporated. The product was purified by rapid chromatography (silica, 25 g silica column, 0-60% ethyl acetate in dichloromethane solution) to give (R)-2-(6-(3-fluoropyrrolidone-1-yl)pyridin-3-yl)-4-oxo-4,6-dihydropyrrolo[3,4-c]pyrazole-5(2H)-carboxylic acid tert-butyl ester, a white solid (200.5 mg, 28%).

[1053] 1 H NMR (400MHz, DMSO-d6) δ = 8.92 (s, 1H), 8.60 (d, 1H), 8.01 (dd, 1H), 6.67 (d, 1H), 5.46 (d, 1H), 4.80(s, 2H), 3.86-3.57(m, 2H), 3.54-3.44(m, 2H), 2.36-2.12(m, 2H), 1.53(s, 9H).

[1054] MS: 388.15 [M+H] +

[1055] Preparation Examples 3-3D

[1056] The following preparation examples were prepared using the halogenated raw materials and suitable amines shown in Table 1a below, following the Pd-coupling method described in Preparation Example 2.

[1057] Table 1a:

[1058]

[1059]

[1060] Preparation Example 4

[1061]

[1062] In a microwave-safe vial, 250 mg (0.659 mmol) of Preparation Example 1 and (S)-pyrrolidine-3-ol (172 mg, 1.978 mmol) were mixed in 10 mL of ethanol. The vial was microwave-safe at 150 °C for 30 min. Then, (S)-pyrrolidine-3-ol (172 mg, 1.978 mmol) was added, and the reaction mixture was microwave-safe at 150 °C for another 45 min. The reaction mixture was filtered and washed with ethanol to give (S)-2-(6-(3-hydroxypyrrolidine-1-yl)pyridin-3-yl)-5,6-dihydropyrrolo[3,4-c]pyrazole-4(2H)-one as a white solid (83.5 mg, 44.4%). 1 H NMR (80MHz, DMSO-d6) δ = 8.60 (s, 1H), 8.51 (d, 1H), 8.07 (s, 1H), 7.92 (dd, 1H), 6.56 (d, 1H), 4.97(d, 1H), 4.34(s, 3H), 3.69-3.37(m, 4H), 2.24-1.80(m, 2H).MS: 286.05[M+H] +

[1063] Each of the following embodiments 5

[1064]

[1065] Preparation Example 2 (160 mg, 0.413 mmol) was stirred in 10 mL of 4 M HCl in dioxane at RT for 3 h 30. The solvent was evaporated under reduced pressure, and the solid was dissolved in dichloromethane. A saturated NaHCO3 solution was added, and the aqueous phase was extracted twice with dichloromethane. The combined organic layers were dried with Na2SO4, filtered, and concentrated to dryness to give (R)-2-(6-(3-fluoropyrrolidone-1-yl)pyridin-3-yl)-5,6-dihydropyrrolo[3,4-c]pyrazole-4(2H)-one, as a white solid (101.5 mg, 86%). 1 H NMR (80MHz, DMSO-d6) δ = 8.62 (s, 1H), 8.55 (d, 1H), 7.96 (dd, 1H), 6.63 (d, 1H), 5 .75(s, 1H), 4.34(s, 2H), 3.92-3.37(m, 4H), 2.45-1.78(m, 2H).MS: 287.80[M+H] +

[1066] Optional preparation example 5

[1067]

[1068] In an argon atmosphere, Preparation Example A (400 mg, 0.1433 mmol), (R)-3-fluoropyrrolidine hydrochloride (720 mg, 5.73 mmol), and cesium fluoride (1306 mg, 8.60 mmol) were mixed in a vial in dry DMSO (4 mL). The reaction mixture was purged with argon and stirred at 120 °C for 6 h 30 min. The reaction mixture was cooled and poured into cold water (pre-cooled in an ice bath). The resulting solution was filtered, and the solid was washed with water. 1 mL of isopropanol was used to directly grind the solid in glass enamel, and the solid was dried to give the product, a light brown solid (287 mg, 0.998 mmol, 70%). 1 H NMR (80MHz, DMSO-d6) δ8.63 (s, 1H), 8.55 (d, 1H), 8.15-7.79 (m, 2H), 6.64 (d, 1H), 5.46(d, IH), 4.34(s, 2H), 3.96-3.40(m, 4H), 2.28-1.56(m, 2H).MS: 288.11[M+H] +

[1069] Optional preparation examples 4-4K

[1070] The following preparation examples were prepared using suitable amines shown in Table 1b below, following the SNAr method described in optional preparation example 5.

[1071] Table 1b:

[1072]

[1073]

[1074]

[1075]

[1076] Preparation Examples 6-6D

[1077] The following preparation examples were prepared according to the deprotection method of Preparation Example 5.

[1078] Table 2:

[1079]

[1080]

[1081] Preparation Example 7

[1082]

[1083] In a vial under an argon atmosphere, a mixture of palladium(II) acetate (13.14 mg, 0.059 mmol) and xantphos (50.8 mg, 0.088 mmol) in 1,4-dioxane (3 mL) was degassed with argon and heated at 100 °C in a preheated block for a few seconds to form a pd-xantphos complex. Then, 83.5 mg (0.293 mmol), 3-iodopyridine (66.0 mg, 0.322 mmol), and cesium carbonate (286 mg, 0.878 mmol) from Preparation Example 4 were added, the mixture was degassed with argon, and heated at 100 °C for 45 min. The reaction mixture was filtered and washed with ethyl acetate. The filtrate was recovered and evaporated to give the product as a yellow, resinous solid (134.5 mg, 0.371 mmol, quantitative).

[1084] 1 H NMR (80MHz, DMSO-d6) δ = 9.04 (d, 1H), 8.83 (s, 1H), 8.57 (d, 1H), 8.43-8.13 (m, 2H), 7.96 (dd, 1H), 7.44 (dd, 1H ), 6.58(d, 1H), 5.08(s, 2H), 4.99(d, 1H), 4.42(d, 1H), 3.64-3.40(m, 4H), 2.17-1.75(m, 2H).MS: 363.08[M+H] +

[1085] Preparation Example 8

[1086] The following preparation examples were prepared using the amide raw materials shown in Table 3 and appropriate halogenated heteroaryl groups according to the Pd-coupling method described in Preparation Example 7.

[1087] Table 3:

[1088]

[1089] Preparation Example 9

[1090]

[1091] Step A

[1092] Under an argon atmosphere, 4,4,5,5-tetramethyl,-2-(prop-1-en-2-yl)-1,3,2-dioxane (0.451 g, 2.7 mmol), [(dppf)PdCl2] (146 mg, 0.179 mmol), and Cs2CO3 (1.16 g, 3.58 mmol) in H2O (0.2 ml) were added to a solution of 2-(6-bromopyridin-3-yl)-5,6-dihydropyrrolo[3,4-c]pyrazole-4(2H)-one (0.5 g, 1.79 mol) in dioxane (20 ml). The mixture was heated at 80 °C for 2 h. The mixture was cooled, and the solvent was evaporated under high vacuum. The residue was dissolved in ethyl acetate, and the solid was filtered off. The residue was washed with water and dried to give 0.450 g of the product. MS: 241.1 [M+H] +

[1093] Step B

[1094] Pd / C (100 mg, 5%) was added to a solution of 2-(6-(prop-1-en-2-yl)pyridin-3-yl)-5,6-dihydropyrrolo[3,4-c]pyrazole-4(2H)-one (1 g, 4.16 mmol) in MeOH (75 mL). The mixture was stirred at room temperature under a H2 (15 psi) atmosphere for 12 hours. Upon completion, the reaction slurry was filtered, and the filtrate was concentrated to give 2-(6-(prop-2-yl)pyridin-3-yl)-5,6-dihydropyrrolo[3,4-c]pyrazole-4(2H)-one (0.85 g). MS: 243.2 [M+H] +

[1095] Preparation Example 10

[1096] The following preparation examples were prepared using suitable borate esters shown in Table 3b, as described in optional preparation example 9.

[1097] Table 3b:

[1098]

[1099] Preparation Example 11

[1100]

[1101] At RT, 50 μL (0.7176 mmol) of 2-propanol in 0.4 mL DMF was added to a suspension of sodium hydride (36 mg / 60% solution in mineral oil, 0.9 mmol) in 2 mL DMF. The mixture was stirred for 30 minutes, and then added at 60 °C to a stirred solution of 2-(6-bromopyridin-3-yl)-5,6-dihydropyrrolo[3,4-c]pyrazole-4(2H)-one (100 mg, 0.358 mmol) in 2 mL DMF. The reaction mixture was heated at 60 °C for 20 hours. After cooling to RT, water and ethyl acetate were added, and the layers were separated. The aqueous layer was extracted with ethyl acetate, the organic layers were combined, dried over MgSO4, filtered, and concentrated under reduced pressure to give 2-(6-isopropoxypyridin-3-yl)-5,6-dihydropyrrolo[3,4-c]pyrazol-4(2H)-one (0.16 g, 35%): MS: 259.2 [M+H] +

[1102] Preparation Example 12

[1103]

[1104] In a sealed test tube under a nitrogen atmosphere, (R)-2-(6-(3-fluoropyrrolidone-1-yl)pyridin-3-yl)-5,6-dihydropyrrolo[3,4-c]pyrazole-4(2H)-one (120 mg, 0.417 mmol), 2-bromo-5-((2-(trimethylsilyl)ethoxy)methoxy)pyridine (253 mg, 0.835 mmol), cuprous iodide (I) (16 mg, 0.0835 mmol), and potassium carbonate (115 mg, 0.835 mmol) were added, and the system was purged with nitrogen. 1,4-Dioxane (6 mL) and N,N'-dimethylethylenediamine (0.017 mL, 0.167 mmol) were added, and the mixture was stirred at 100 °C for 4 h. The reaction mixture was concentrated under reduced pressure, and the residue was dissolved in 10 ml of water and extracted with DCM / MeOH (9:1, 50 ml x 2). The combined organic layers were dried over NaSO4 (5 g), filtered, and concentrated to give 80 mg of a pale yellow solid crude product. The crude product was purified by alkaline silica gel (100-200 mesh) column chromatography using a dichloromethane / methanol gradient (100 / 0->98 / 2) to give the desired product as a pale yellow solid (50 mg, 23% yield). 1H NMR (400MHz, DMSO-d6) δ8.86 (s, 1H), 8.60 (d, 1H), 8.42-8.33 (m, 1H), 8.18 (dd, 1H), 8.01 (dd, 1H), 7.58 (dd, 1H), 6.66 (d, 1H), 5.54 (s , 1H), 5.28(s, 2H), 5.05(s, 2H), 3.85-3.54(m, 5H), 3.54-3.42(m, 1H), 2.39-2.08(m, 2H), 0.90(dd, 2H), -0.01(s, 9H).MS: 511.3[M+H] +

[1105] Preparation Examples 13-31

[1106] The following preparation examples were prepared using the amide raw materials shown in Table 3c and appropriate haloaryl groups according to the Cu-coupling method described in Preparation Example 12.

[1107] Table 3c:

[1108]

[1109]

[1110]

[1111]

[1112]

[1113]

[1114]

[1115] Examples 1-4

[1116] The following compounds were prepared using the amide starting materials shown in Table 4 and appropriate halogenated heteroaryl groups according to the Pd-coupling method described in Preparation Example 7.

[1117] Table 4:

[1118]

[1119]

[1120] Alternative Example 1

[1121]

[1122] In a flask under an argon atmosphere, the following ingredients were mixed: 285 mg (0.992 mmol) of Preparation Example 5, 3-bromopyridine (0.191 mL, 1.984 mmol), potassium carbonate (274 mg, 1.984 mmol), and cuprous iodide (I) (37.8 mg, 0.198 mmol). The system was purged with argon. Dioxane (12 mL) and N1,N2-dimethylethyl-1,2-diamine (0.042 mL, 0.397 mmol) were added, and the mixture was stirred at 110 °C for 4 h. The crude product was concentrated under reduced pressure and dissolved in 20 mL of water. Ammonia (16.30 mL, 114 mmol) was added until the solution was alkaline (pH 12). The aqueous layer was extracted twice with DCM / MeOH solution (9:1). The combined organic layers were dried over Na2SO4, filtered, and concentrated to dryness. The solids were suspended in DCM and stirred at 40 °C for 15 min. The mixture was cooled and filtered to give the product as a white solid (234.3 mg, 65%). 1 H NMR (80MHz, DMSO-d6) δ9.03 (d, 1H), 8.86 (s, 1H), 8.60 (d, 1H), 8.42-8.15 (m, 2H), 8.01 (dd, 1H), 7.45 ( dd, 1H), 6.67 (d, 1H), 5.41 (d, 1H), 5.09 (s, 2H), 4.00-3.37 (m, 4H), 2.28-1.48 (m, 2H).MS: 365.12[M+H] +

[1123] Example 5-138

[1124] The following examples were prepared according to the method described in Preparation Example 7 or Alternative Example 1, or using the amide starting materials shown in Table 4a below and appropriate haloaryl groups. Alternatively, Pd2(dba)3, BINAP, and Cs2CO3 conditions can be applied.

[1125] Table 4a:

[1126]

[1127]

[1128]

[1129]

[1130]

[1131]

[1132]

[1133]

[1134]

[1135]

[1136]

[1137]

[1138]

[1139]

[1140]

[1141]

[1142]

[1143]

[1144]

[1145]

[1146]

[1147]

[1148]

[1149]

[1150]

[1151]

[1152]

[1153]

[1154]

[1155]

[1156]

[1157]

[1158]

[1159]

[1160] Example 139

[1161]

[1162] A suspension of 2-(6-(pyrrolidone-1-yl)pyridin-3-yl)-5,6-dihydropyrrolo[3,4-c]pyrazole-4(2H)-one (0.08 g, 0.286 mmol), 2,6-difluoropyrazine (0.199 g, 1.72 mmol), and CsF (0.348 g, 2.293 mmol) in DMSO (4 mL) was heated at 130 °C for 30 min under microwave irradiation. The reaction mixture was then cooled and poured into ice-cold water (3 mL). The resulting slurry was filtered, and the solids were washed with water (5 mL). The residue was purified by silica gel (100-200 mesh) column chromatography using a 2-5% MeOH solution in DCM to give the desired product (32 mg, 29%). 1 H-NMR (400MHz, DMSO-d6) δ9.65-9.59 (m, 1H), 8.96 (s, IH), 8.60-8.55 (m, 1H), 8.38 (dd, IH), 7 .97(dd, 1H), 6.59(d, 1H), 5.03(s, 2H), 3.46-3.41(m, 4H), 2.01-1.93(m, 4H).MS: 366.1[M+H] +

[1163] Examples 140-161

[1164] The following examples were prepared using the amide raw materials shown in Table 4b and suitable amides and fluoro-heteroaryl groups, as described in Example 54.

[1165] Table 4b:

[1166]

[1167]

[1168]

[1169]

[1170]

[1171]

[1172] Example 162

[1173]

[1174] (R)-2-(6-(3-fluoropyrrolidine-1-yl)pyridin-3-yl)-5-(5-(((2-(trimethylsilyl)ethoxy)methoxy)pyridin-2-yl)-5,6-dihydropyrrolo[3,4-c]pyrazole-4(2H)-one (50 mg, 0.098 mmol) was dissolved in DCM (1.5 mL) and cooled to 0 °C in an ice bath with stirring. A solution of 1,4-dioxane in 4 M HCl (0.2 mL) was added to this solution, and the mixture was stirred continuously at RT for 4 h. After the reaction was complete, the solvent was removed under reduced pressure, and the solution was alkalized to pH 8-9 with a saturated sodium bicarbonate aqueous solution. The compound was precipitated, and the solid was collected by filtration. The solid was washed with pentane (3 mL) and dried under high vacuum for 30 min to give the desired compound as a pale yellow solid (10 mg, 27%). 1 H NMR (500MHz, CF3COOD) δ8.85 (s, 1H), 8.72 (d, 1H), 8.57 (dd, 1H), 8.52-8.32 (m, 2H), 7.90 (d, 1H), 7.46 (s , 1H), 5.86-5.62(m, 1H), 5.53(s, 2H), 4.52-4.01(m, 4H), 2.87(s, 1H), 2.75-2.44(m, 1H).MS: 381.1[M+H] +

[1175] Examples 163-181

[1176] The following examples were prepared using the O-protected raw materials shown in Table 4c, following the deprotection method shown in Example 162.

[1177] Table 4c:

[1178]

[1179]

[1180]

[1181]

[1182]

[1183] Precursor 1

[1184]

[1185] In a flask under an argon atmosphere, the preparation of Example 7 (135 mg, 0.373 mmol) was dissolved in dichloromethane. Triethylamine (1.038 mL, 7.45 mmol) was added, and the reaction mixture was stirred for 5 minutes. Then, methanesulfonyl chloride (0.290 mL, 3.73 mmol) was added dropwise to the reaction mixture. The mixture was stirred at room temperature for 20 minutes. Methanesulfonyl chloride (0.290 mL, 3.73 mmol) was added, and the reaction mixture was stirred for 25 minutes. The reaction mixture was quenched with 1 N NaOH aqueous solution, and then extracted three times with dichloromethane. The combined organic layers were dried over Na2SO4, filtered, and concentrated to dryness. The product was purified by rapid chromatography (silica, 12 g silica column; 0-10% methanol in dichloromethane solution) to give (S)-methanesulfonic acid 1-(5-(4-oxo-5-(pyridin-3-yl)-5,6-dihydropyrrolo[3,4-c]pyrazol-2(4H)-yl)pyridin-2-yl)pyrrolidine-3-yl ester, as a white solid (17.4 mg, 11%). 1 H NMR (80MHz, DMSO-d6) δ = 9.03 (d, 1H), 8.87 (s, 1H), 8.61 (d, 1H), 8.44-8.15 (m, 2H), 8.03 (dd, 1H), 7.45 (q, 1H), 6.68(d, 1H), 5.45(s, 1H), 5.09(s, 2H), 3.67(d, 4H), 3.27(s, 3H), 2.41-2.08(m, 2H).MS: 441.08[M+H] +

[1186] Alternative methods

[1187] In a vial cooled to 0°C under an argon atmosphere, (S)-2-(6-(3-hydroxypyrrolidone-1-yl)pyridin-3-yl)-5-(pyridin-3-yl)-5,6-dihydropyrrolo[3,4-c]pyrazole-4(2H)-one (100 mg, 0.276 mmol) and 4-dimethylaminopyridine (337 mg, 2.76 mmol) were mixed in pyridine (17 mL). Methanesulfonyl chloride (0.108 mL, 1.380 mmol) was added, and the mixture was purged with argon. The reaction mixture was warmed to RT and stirred for 2 h, after which 4-dimethylaminopyridine (169 mg, 1.380 mmol) and methanesulfonyl chloride (0.054 mL, 0.690 mmol) were added at 0°C. After 40 min, 0.1 N NaOH aqueous solution (20 mL) was added to the mixture to alkalize it. Pour the solution into cold water and filter. Wash with water until the pH of the water reaches 7. Dry the solid under high vacuum for 30 min to obtain the compound as an orange solid (86 mg, 71%). 1H NMR (400MHz, DMSO-d6) δ9.03 (d, 1H), 8.87 (s, 1H), 8.61 (d, 1H), 8.35 (d, 1H), 8.26 (d, 1H), 8.02 (dd, 1H), 7.45 (dd , 1H), 6.68(d, 1H), 5.44(s, 1H), 5.08(s, 2H), 3.86-3.42(m, 4H), 3.27(s, 3H), 2.40-2.24(m, 2H).MS: 441.1[M+H] +

[1188] Precursor 2

[1189]

[1190] N-bromosuccinimide (22 mg, 0.126 mmol) was added to a solution of Preparation Example 8 (43 mg, 0.097 mmol) in dimethylformamide (3 mL). After stirring at room temperature for 1 h, the reaction mixture was diluted with water and ethyl acetate. The layers were separated, and the aqueous layer was extracted twice with ethyl acetate. The combined organic layers were dried over sodium sulfate, filtered, and concentrated under reduced pressure. The residue was ground in acetonitrile, and the solid was collected by filtration. The crude solid was then purified by rapid chromatography (silica, 12 g silica column; 2-5% methanol in dichloromethane solution). The fraction was concentrated under reduced pressure, and the residue was ground in acetonitrile. The solid was collected by filtration to obtain (R)-2-(5-bromo-6-(3-fluoropyrrolidone-1-yl)pyridin-3-yl)-5-(5-bromopyridin-3-yl)-5,6-dihydropyrrolo[3,4-c]pyrazole-4(2H)=one, a light brown solid (16 mg, 32%).

[1191] 1 H-NMRδ8.88(d,1H),8.83-8.70(m,1H),8.51-8.32(m,2H),8.22-7.83(m,2H ), 5.75-4.77(m, 3H), 4.32-3.72(m, 4H), 0.98-0.67(m, 2H).MS: 523.10[M+H] +

[1192] Precursor 3

[1193]

[1194] Triethylamine (0.08 mL, 0.5797 mmol) was added to a solution of Preparation Example 7 (70 mg, 0.193 mmol) in DCM (3.5 mL) under a nitrogen atmosphere at RT. The reaction mixture was cooled to 0 °C, and then p-toluenesulfonyl chloride (73 mg, 0.3865 mmol) was added incrementally over a 10-minute period, followed by the addition of DMAP (23 mg, 0.193 mmol). The reaction mixture was then warmed to RT and stirred for 12 h, with the reaction progress monitored by TLC. After the reaction was complete, the mixture was diluted at RT with a saturated aqueous solution of NaHCO3 (5 mL) and extracted twice (2 x 20 mL) with a 5% MeOH solution in DCM. The combined organic layers were dried over Na2SO4. The solvent was distilled off under reduced pressure to give a pale yellow solid. The crude compound was purified by alkaline silica gel (100-200 mesh) column chromatography, eluting with a DCM / MeOH gradient (100 / 0->98 / 2) to obtain the desired compound as a yellowish-white solid (20 mg, 20%). 1 H NMR (400MHz, DMSO-D6) δ9.03 (d, IH), 8.86 (d, 1H), 8.58 (dd, 1H), 8.35 (dd, 1H), 8.32-8.18 (m, 1H), 7.99 (ddd, 1H), 7.69-7 .54(m, 2H), 7.44(td, 3H), 6.62(dd, 1H), 5.25-5.01(m, 3H), 3.76-3.39(m, 4H), 2.39(d, 3H), 2.36-2.01(m, 2H).MS: [M+H] + 517.3

[1195] Precursor 4

[1196]

[1197] In an argon atmosphere, 700 mg (1.93 mmol) of Preparation Example 7 and 4-DMAP (236 mg, 1.93 mmol) were suspended in 9.4 mL of pyridine in a flask and cooled to 0 °C. 2.14 g (9.66 mmol) of 4-nitrobenzenesulfonyl chloride was added, and the suspension was stirred at room temperature for 4 h. 118 mg (0.97 mmol) of 4-DMAP and 1.07 g (4.83 mmol) of 4-nitrobenzenesulfonyl chloride were added at 0 °C. The reaction mixture was stirred overnight. 118 mg (0.97 mmol) of 4-DMAP and 1.07 g (4.83 mmol) of 4-nitrobenzenesulfonyl chloride were added again at 0 °C, and the reaction mixture was stirred at room temperature for 1 day. 40 mL of 1 M NaOH was added, and the resulting mixture was centrifuged at 6000 ppm for 5 min. The mixture was decanted into a centrifuge vial, and the remaining solid was washed four times with 40 mL of water. Water was removed by centrifugation / decantation after each washing step. The remaining solid was suspended in water, transferred to a flask, and evaporated to give the desired product, a light brown solid (871 mg, 83%). 1 H NMR (400MHz, DMSO-d6) δ = 9.04 (s, 1H), 8.85 (s, 1H), 8.68-7.90 (m, 8H), 7.46 (bs, 1H), 6 .63(d,1H),5.42(s,1H),5.09(s,2H),3.86-3.39(m,4H),2.36-2.05(m,2H).MS:[M+H] + 547.97

[1198] Radioligand synthesis

[1199] Example 1 3 H-1]

[1200]

[1201] In a tritium reaction vessel, precursor 2 (0.5 mg) was dissolved in dimethylformamide (DMF) (0.3 mL) and N,N-diisopropylethylamine (DIEA) (5 μL). 10% Pd / C (0.5 mg) was added, and the vessel was pressurized to 0.5 atm with tritium gas at -200 °C. The solution was stirred at room temperature for 1 h, cooled to -200 °C, and excess gas was removed. The reaction flask was rinsed with 4 x 1 mL CH3OH, ensuring each CH3OH rinse passed through the diatomaceous earth layer. Combined methanol was removed under vacuum. The substance was purified by HPLC. The mobile phase was removed, and the product was redissolved in anhydrous ethanol. (5 mCi, radiochemical purity >99%, specific activity 43.6 Ci / mmol). T refers to tritium ( 3 H). MS (ESI): m / z=369 (100%) [M+H ] +

[1202] Example 1-[ 18 F-1]

[1203]

[1204] Drying step: In a typical method, the [[] in the transport vials 18 F] Fluoride (target water obtained from a commercial cyclotron accelerator facility) is transferred to and captured on an ion exchange column. It is then eluted with a solution of potassium carbonate and Kryptofix 222. The reaction vessel (RV1) of the module was used. The solution was first evaporated by heating at 95°C for 4 min under vacuum and a helium atmosphere. Acetonitrile (1 mL) was added to RV1, and evaporation continued for 2 min under the same conditions under vacuum and a helium atmosphere. After a second addition of acetonitrile (1 mL), final evaporation was carried out at 95°C for 2 min under vacuum and a helium atmosphere. The reactor was then cooled to 60°C.

[1205] Radiolabeling: A solution of precursor 1 (1 mg) in anhydrous dimethyl sulfoxide (0.7 mL) was added to the reaction vessel, and the reaction mixture was heated at 100 °C for 10 min. The reactor was cooled to 40 °C, diluted with HPLC mobile phase (1.8 mL), and the contents were transferred to a loop loading vial (RV2). The reactor was rinsed with water for injection (2.5 mL), and the rinse solution was transferred to RV2. The contents of RV2 were transferred to an HPLC syringe loop for purification.

[1206] HPLC Purification: Purification was performed by HPLC using a semi-preparative Phenomenex Synergi C18 column (5 μm, 250 x 10 mm), eluting with a mixture of acetonitrile / ammonium acetate (20 mM) (35 / 65, v / v) at a flow rate of 4 mL / min. The product fraction was collected in flask 1 containing 20 mL of sodium ascorbate (5 mg / mL) in WFI. The diluted product mixture was passed through a C18 solid-phase extraction column, and the column was washed with 10 mL of WFI solution containing sodium ascorbate (5 mg / mL). The radiolabeled product was eluted from the SPE column to a formulation flask pre-filled with 10 mL of formulation base (a saline solution of sodium ascorbate (4.67 mg / mL)) using 1.0 mL of 200 standard strength USP grade ethanol. The column was washed with 4.0 mL of formulation base, and the wash solution was mixed with the contents of the formulation flask. The resulting solution was passed through a sterile 0.2 μm membrane filter into a sterile, filter-connected vial (final product vial, FPV), which was pre-filled with 15 mL of physiological saline (27% attenuation-corrected yield).

[1207] Example 4 3 H-4]

[1208]

[1209] Example 4 (1.0 mg) was added to a tritium reaction vessel, followed by cesium carbonate (1.0 mg), then DMF (0.1 mL), and finally iodomethane [3H] (100 mCi). The vessel was sealed and the solution was stirred at room temperature for 18 h. The reaction mixture was transferred to a larger flask. The reaction vessel was rinsed with 4 x 2 mL methanol. The combined methanol was removed under vacuum. Crude yield: 38 mCi. The substance was purified by silica gel column chromatography. The mobile phase was removed under vacuum, and the product was redissolved in a 0.05% TFA solution in water / acetonitrile. The substance was then purified by semi-preparative reversed-phase HPLC. The mobile phase was removed under vacuum, and the product was redissolved in anhydrous ethanol (4.8 mCi, purity >99%). The specific activity was determined to be 79.98 Ci / mmol by MS.

[1210] MS(ESI): m / z=374(100%)[M+H] +

[1211] Biological assay description and corresponding results

[1212] 1. Preparation of α-synuclein aggregates derived from the brain of human Parkinson's disease (PD) patients.

[1213] This method is adapted from the protocol described by Spillantini et al., 1998. Frozen tissue blocks from PD donors were thawed on ice and homogenized using a glass Doens homogenizer. The homogenate was then centrifuged for 20 minutes at 11,000 x g (12,700 RPM) at 4 °C using a pre-chilled 70.1 rotor (Beckman, 342184). The precipitate was resuspended in extraction buffer [10 mM Tris-HCl pH 7.4, 10% sucrose, 0.85 mM NaCl, 1% protease inhibitor (Calbiochem 539131), 1 mM EGTA, 1% phosphatase inhibitor (Sigma P5726 and P0044)] and centrifuged for 20 minutes at 15,000 x g (14,800 rpm, 70.1 Ti rotor) at 4 °C. Discard the precipitate and add sodium dodecyl sarcosinate (20% stock solution, Sigma L7414) to the supernatant to a final concentration of 1% at room temperature, and hold for 1 hour. Then centrifuge the solution at 100,000 × g (38,000 rpm, 70.1 Ti rotor) at 4 °C for 1 hour. Resuspend the precipitate containing enriched α-syn aggregates in PBS and store at -80 °C until use.

[1214] 2. Microscopic radioactive binding competition assay for determining binding affinity

[1215] PD-derived α-syn aggregates were spotted onto a microarray slide. The slide was then coated with a 20 nM tritium-modified reference ligand. 3 H]-a-syn-Ref (as described in WO2017 / 153601) and the compound of the present invention (non-radiolabeled) in increments of 1 mM or 50 pM-2 μM were incubated together. After incubation, the slide was washed and exposed to a phosphor storage screen (GE Healthcare, BAS-IP TR 2025). After exposure, the phosphor storage screen was scanned with a laser imaging system (Typhoon FLA 7000) to read out the signal from the above-described radiobinding experiment. The signal was quantified using the ImageJ software package. Non-specific signals were measured with an excess of the non-radiolabeled reference ligand (1 mM), and specific binding was calculated by subtracting the non-specific signal from the total signal. Competition was calculated as a percentage, where 0% was defined as specific binding in the presence of the solvent, and 100% was defined as the value obtained in the presence of an excess of the non-radiolabeled reference ligand. All measurements were performed at least twice technically. K was calculated in GraphPad Prism7 by applying a nonlinear regression curve fitting using a unit point specific binding model. i value.

[1216] The potency of the example compounds in competitive binding to α-syn aggregates derived from the brains of PD patients was evaluated against a [3H] radiolabeled reference ligand. Results of the micro-radiobinding competition assays of the example compounds tested are shown in Table 5 as competition % at 1 μM and Ki values. All measurements were performed on the same PD brain-derived α-syn aggregates. The Ki value for compound 1 reported herein is the average of two independent experiments.

[1217] Table 5:

[1218]

[1219] Table 5: Evaluation of binding affinity by microradioactive binding competition assay of α-syn aggregates derived from human PD brain. Left: Percentage (%) of competition with the tritized reference ligand in the presence of 1 μM of Example Compounds 1 and 2. Right: Ki value of Example Compound 1 is shown. As shown in Table 5, Example Compounds 1 and 2 of the present invention exhibit good binding to α-syn aggregates derived from PD brain.

[1220] 3. Example 1 [ 3 Evaluation of target binding of H-1 in α-synucleinopathia and AD tissues

[1221] 3A: Through high-resolution microautoradiography

[1222] This protocol is a modification of Marquie et al., 2015. The slides were incubated with 60 nM tritized compound 1 of the examples (Example-1 [3H-1]) or the reference ([3H]-Tau-Refat 60 nM) at room temperature (RT) for 1 hour. The slides were then washed as follows: once in ice-cold 50 mM Tris-HCl pH 7.4 buffer for 1 minute each time, twice in ice-cold 70% ethanol for 1 minute each time, once in ice-cold 50 mM Tris-HCl pH 7.4 buffer, and finally simply rinsed in ice-cold distilled water. The slides were then dried and exposed to Ilford NuclearEmulsion Type K5 (Agar Scientific, AGP9281) in a light-protected slide storage box. Five days later, the sections were developed by continuously immersing them in the following solutions: 1.) Ilford Phenisol developer (1:5 diluted in H2O, Agar Scientific, AGP9106), 2.) IlfoStop solution (1:20 diluted in H2O, Agar Scientific, AGP9104), 3.) Ilford Hypam fixer (1:5 diluted in H2O, Agar Scientific, AGP9183), and finally with H2O. z O Rinse.

[1223] Immunostaining was also performed on the same sections during display. For image acquisition, sections were fixed using ProLong Gold anti-fading reagent (Invitrogen P36930) and imaged on a Panoramic 150 slide scanner (3DHiStech) with 20x objectives, capturing bright-field and fluorescence images respectively.

[1224] 3B. Staining sections using antibodies

[1225] Immunostaining of brain sections was performed using a commercially available antibody specific to phosphorylated serine at amino acid 129 (α-syn-pS129, rabbit monoclonal, Abcam 51253), a mouse conformation-dependent anti-Tau antibody (MC1, courtesy of Peter Davies, Northwell, US), or a commercially available antibody specific to TDP-43 phosphorylated serine at amino acids 409 / 410 (anti-PTDP-43pS409 / 410, Biolegend 829901). Sections were fixed at 4°C with 4% formaldehyde (Sigma, 252549) for 15 minutes and washed three times with 1×PBS (Dulbecco phosphate-buffered saline, Sigma D1408) for 5 minutes each time. Next, the slides were saturated and permeabilized in blocking buffer (PBS, 10% NGS, 0.25% Triton X-100) at room temperature for 1 hour, and then incubated overnight at 4°C with primary antibodies corresponding to α-syn-pS129 or MC1 (PBS, 5% NGS, 0.25% Triton X-100). The next day, the slides were washed three times with 1x PBS for 5 minutes each time, and then incubated with Alexa Fluor 647-labeled goat anti-rabbit (Abcam, ab150079) or goat anti-mouse (115-605-166, Jackson ImmunoResearch) secondary antibodies at room temperature for 45 minutes. After incubation with the secondary antibodies, the slides were washed three times in PBS and then processed further. For image acquisition, slides were mounted using ProLong Gold anti-fading reagent (Invitrogen P36930) and imaged using a Panoramic 150 slide scanner (3DHIstech; Hungary).

[1226] result Example 1: Frozen human brain sections from cases of different α-synucleinosis [ 3 High-resolution micro-autoradiography of H-1]. Detection of H-1 in PD and other α-synucleinopathies (including multiple system atrophy (MSA), Lewy body dementia (DLB), Lewy body variant of Alzheimer's disease (LBV), and PDD). 3 The strong autoradiographic signal of H-1] Figure 1 (Lower part), and co-localized with the immunofluorescence signal from the α-syn-pS129 antibody ( Figure 1 (Upper part), which indicates strong target binding on the Lewy body and Lewy neurites, as well as on very small α-syn aggregates.

[1227] 4. Example 1: Autoradiography of brain slices from PD, PDD, and non-dementia control (NDC) donors [ 3 Assessment of the specific binding of H-1]

[1228] Frozen human brain sections from one case of familial PD (α-synuclein [SNCA] gene G51D missense mutation) (labeled SNCA(G51D)), one case of PDD, and two non-dementia controls (NDC) were first briefly fixed at 4°C with 4% paraformaldehyde (Sigma, 252549) for 15 minutes, and washed three times with PBS (Dulbecco phosphate-buffered saline, Sigma) for 5 minutes each time at RT. All slides were then equilibrated in 50 mM Tris-HCl pH 7.4 buffer for 20 minutes before use in experiments. Each brain section was then subjected to a fixed concentration (10 nM) of tritium-modified compound 1 of Example 1 (Example-1 […]) at RT. 3 Example 1: H-1] or reference α-syn ligand ([3H]-α-syn-ref) or increasing concentrations in the range of 1.25 nM to 80 nM. 3 The tritium compound of [H-1] was incubated together in Tris-HCl buffer for 2 hours (total binding, '-'). To determine nonspecific (NS) binding, the tritium compound of Example-1 [ 3 [H-1] or [3H]-a-syn-Ref was mixed with 1 μM of a non-radioactively labeled compound (for example or a-syn-Ref, self-blocking, '+', respectively). The slide was washed and placed under a phosphor imaging screen (GE Healthcare, BAS-IP TR 2025) in the imaging cassette. The imaging screen was scanned using a laser imaging system (Typhoon, FLA 7000), and the resulting images were analyzed using the ImageJ software package. Specific binding was determined by subtracting the non-specific signal from the total signal. d The values ​​are calculated in GraphPad Prism7 by applying a nonlinear regression curve fit using a unit-point specificity combined with the model.

[1229] result: Example 1 3 [H-1] showed dose-dependent autoradiographic signals in various α-synucleinopathy tissues, including PDD ( Figure 2A ) and hereditary PD cases ( Figure 3A In both cases, the alternative signals correlated well with the localization of α-syn pathology, as determined by staining with α-syn-pS129 antibody, indicating specific binding of the compound to PDD and PD tissues. Figure 2B and 3BThe dissociation constant (Ki) was calculated at 11–13 nM using a quantitative specific signal. d ()( Figure 2C Table 6 and Figure 3C (Table 6) shows a good binding affinity for pathological α-synuclein aggregates.

[1230] Table 6:

[1231]

[1232] Table 6: Examples of Autoradiography-1 [ 3 [H-1] Assessment of binding affinity from human brain tissue of idiopathic PD cases (PDD) and familial PD cases (G51D missense mutation). The dissociation constant (K) was calculated by applying nonlinear regression curve fitting using a single-point specific binding model in GraphPad Prism7. d ) and binding site occupancy (B maax R 2 It is the coefficient of determination.

[1233] Additionally, when compared to the reference α-syn ligand, Example-1 [ 3 [H-1] showed improved overall binding and excellent specific binding in tissues from various α-synucleinosis cases, and extremely weak binding in disease-free tissues (NDC). Figure 4A and Figure 4B ).

[1234] 5. Saturation binding study of PD brain-derived α-syn aggregates using microradiotherapy

[1235] PD-derived α-syn aggregates were spotted onto a microarray slide. The slide was then compared with that of Example-1. 3 After incubation with [H-1] or [3H]-α-syn-ref (concentration increasing in the range of 300 pM-150 nM), the slides were washed and exposed to a phosphor storage screen (GE Healthcare, BAS-IP TR 2025). Following exposure, the phosphor storage screen was scanned using a laser imaging system (Typhoon FLA7000) to read out the signals from the above radiobinding experiments. Signal quantification was performed using the ImageJ software package. Non-specific signals were measured with excess non-radiolabeled reference ligands (2 μM of Example 1 or α-syn-ref, respectively), and specific binding was calculated by subtracting the non-specific signal from the total signal. K was calculated in GraphPad Prism7 using a unit-point specific binding model with nonlinear regression curve fitting. d value.

[1236] result: Example-1 was evaluated in a study of saturated binding of PD tissue homogenates by microradioactive binding. 3 [H-1], and compared head-to-head with a reference α-syn binder. For example... Figure 5 As shown, Example-1 [ 3 H-1] showed high and improved binding site occupancy on PD brain-derived α-syn aggregates.

[1237] 6. Example 1: Replacement of α-syn-Ref with PD-derived α-syn aggregates via microradiation [ 3 H-1] assessment

[1238] PD-derived α-syn aggregates were spotted onto a microarray slide. The slide was then compared with 20 nM α-syn aggregates from Example-1. 3 [H-1] was incubated with α-syn-Ref or the non-radiolabeled compound of Example 1 at concentrations increasing in the range of 50 pM to 2 μM. After incubation, the slide was washed and exposed to a phosphor storage screen (GE Healthcare, BAS-IP TR 2025). Following exposure, the phosphor storage screen was scanned using a laser imaging system (Typhoon FLA 7000) to read out the signal from the above-described radiobinding experiment. The signal was quantified using the ImageJ software package. Non-specific signals were measured with an excess of the non-radiolabeled compound 1 of Example 1 (2 μM), and specific binding was calculated by subtracting the non-specific signal from the total signal. Competition was calculated as a percentage, where 0% was defined as specific binding in the presence of the solvent, and 100% was defined as the value obtained in the presence of an excess of the non-radiolabeled reference ligand. All measurements were performed in at least two technical replicates.

[1239] result: Evaluation Example 1 [ 3 Can H-1 be replaced by a non-radioactively labeled α-syn-Ref compound? The α-syn-Ref compound is only present on α-syn aggregates derived from the brain of idiopathic PD cases, as described in Example-1. 3 H-1] Partial Competition ( Figure 6 This indicates that, compared to α-syn-Ref compounds, Compound 1 of Example binds to different or partially overlapping binding bags of pathological α-syn aggregates.

[1240] 7. Radioactive binding competition assay for determining the inhibitory constant (Ki) of compound 1 in AD brain homogenate.

[1241] Preparation of human Alzheimer's disease (AD) brain homogenate:

[1242] This method was modified from the protocol described by Bagchi et al., 2013. Frozen tissue blocks from AD donors were thawed on ice and homogenized at 4°C using a glass Doens homogenizer in a high-salt buffer (50 mM Tris-HCl pH 7.5, 0.75 M NaCl, 5 mM EDTA) supplemented with a protease inhibitor (Complete; Roche 11697498001). The homogenate was centrifuged at 100,000 × g (38,000 rpm) for 1 hour at 4°C using a pre-chilled 70.1 rotor (Beckman, 342184). The precipitate was resuspended in a high-salt buffer supplemented with 1% Triton X-100 and homogenized at 4°C using a glass Doens homogenizer. The homogenate was centrifuged again at 100,000 × g (38,000 rpm, 70.1 rotor) for 1 hour at 4 °C. The precipitate was resuspended in high-salt buffer supplemented with 1% Triton X-100 and 1M sucrose and homogenized using a glass Dunns homogenizer at 4 °C. The homogenate was centrifuged again at 100,000 × g (38,000 rpm, 70.1 rotor) for 1 hour at 4 °C. The resulting precipitate containing the insoluble fraction was resuspended in PBS, aliquoted, and stored at -80 °C until use.

[1243] A fixed concentration of the AD insoluble fraction was compared with a 10 nM tritized reference Aβ ligand ([ 3 Compound 1 of Example, containing non-radiolabeled ligands in the range of 400 pM to 2 μM (H]-Aβ-ref), was incubated at RT for 2 hours. The sample was then vacuum filtered through a GF / C filter plate (PerkinElmer) to capture aggregates with bound radioligands and washed five times with 50 mM Tris pH 7.5. The GF / C filter was then dried, and scintillation fluid (Ultimate Gold, PerkinElmer) was added to each well. The filter was analyzed on a MicroBeta2 scintillation counter (PerkinElmer). Non-specific signals were measured with an excess of the non-radiolabeled reference ligand (2 mM), and specific binding was calculated by subtracting the non-specific signal from the total signal. Competition was calculated as a percentage, where 0% was defined as specific binding in the presence of the solvent, and 100% was defined as the value obtained in the presence of an excess of the non-radiolabeled reference ligand. Ki values ​​were calculated in GraphPad Prism 7 by applying a nonlinear regression curve fit using a unit-point specific binding model. Measurements were repeated at least twice.

[1244] result :like Figure 7As shown in Table 7, the Ki value of Compound 1 from Example 1 in the AD brain-derived homogenate was determined at 330 nm. Based on Example-1 [ 3 H-1] showed good selectivity for α-syn compared to pathological aggregates of Aβ present in human AD brain homogenates, exhibiting superior binding affinity for PD brain tissue (via autoradiography) and binding affinity for PD brain homogenates (via microradiographic binding). Furthermore, compared to the reference Tau binder used as a positive control, Compound-1 of Example [ 3 H-1] did not show specific target binding to Tau aggregates in AD brain tissue (Figure 8), indicating good selectivity compared to pathological Tau aggregates. Furthermore, Example-1 [ 3 H-1] showed very weak to no binding to TAR DNA-binding protein 43 (TDP-43) aggregates present in type C brain tissue of frontotemporal degenerative TdP (FTLD-TdP). Figure 9 The results indicate good selectivity compared to TDP-43 pathological aggregates. In summary, these data demonstrate the selectivity of Compound 1 of Example.

[1245] Table 7:

[1246]

[1247] Table 7: Ki values ​​of Compound 1 in AD brain-derived homogenates were determined by replacing [3H]-Abeta-Ref with the non-radioactively labeled Compound 1. Ki was calculated using a unit-point specific binding model in GraphPad Prism 7 and by applying nonlinear regression curve fitting. i and R 2 value.

[1248] 8. PK study in healthy monkeys

[1249] Using 1 ml of ethanol and 14 ml of ascorbate / saline (ascorbate solution prepared at a concentration of 9.3 mg / ml), 18 Example 1 of F-marking [ 18 F-1] (6.5 mCi) intravenous (iv) injection into non-human primates (NHP). Monkey PET scans were performed using a Siemens Focus 220. PET acquisition began immediately before the injection of the radioactive dose. Images were generated as a 120-minute dynamic scan with head-focused imaging. Example-1 [ 18 F-1] has a rapid uptake of 2.0 SUVmax into the whole brain (3.5 min after injection). Furthermore, Example-1 [ 18 F-1] exhibits rapid peak-to-half-peak clearance within 14 minutes. Figure 10This data proves the validity of Example 1. 18 The PK curve of F-1 in non-human primates is suitable for its use as a brain PET agent in humans.

[1250] 9. Evaluation of the specific binding of Example-1 [3H-1] in brain slices from PD, PDD, MSA, LBV and non-dementia control (NDC) donors by autoradiography

[1251] First, frozen human brain sections from one PD case, two PDD cases, two MSA cases, one LBV case, and three non-dementia control (NDC) cases were briefly fixed at 4°C with 4% paraformaldehyde (Sigma, 252549) for 15 minutes and washed three times with PBS (Dulbecco phosphate-buffered saline, Sigma) for 5 minutes each time. Then, before use in experiments, all slides were equilibrated in 50 mM Tris-HCl pH 7.4 buffer for 20 minutes. Each brain section was then treated with a fixed concentration (10 nM) of tritium compound 1 from Example 1 (Example-1) in Tris-HCl buffer. 3 H-1]) was incubated at RT for 2 hours (total binding, "total"). To determine nonspecific binding (NSB), Example-1 [ 3 [H-1] was mixed with 5 mM of the non-radioactively labeled compound from Example 1. The slide was washed and then exposed and scanned in a real-time autoradiography system (Beaquant Instrument, AI4R).

[1252] result: Example 1 3 H-1] showed target binding in various α-synucleinopathy tissues, including two MSA, one LBV, and two PDD cases. Figure 11A Displaceable signals correlated well with the localization and load of α-syn pathology, as determined by staining with α-syn-pS129 antibody. Figure 11B The results indicate specific binding of the compound. Furthermore, the autoradiographic signal in the diseased donor was significantly stronger compared to several non-dementia control cases with weak signals.

[1253] 10. Microscopic radioactive binding competition assay for determining binding affinity

[1254] PD-derived α-syn aggregates were spotted onto a microarray slide. The slide was then compared with 6 nM or 20 nM of the sample from Example-1. 3[H-1] was incubated with the example compound (non-radiolabeled) at 1 μM and 100 nM. In some cases, different concentration ranges of the non-radiolabeled example compound were further evaluated, varying from 0.05 nM to 2 μM. After incubation, the slides were washed and scanned using a real-time autoradiography system (Beaquant, AI4R). The signal was quantified using Beamage image analysis software (AI4R). Non-specific signals were determined with an excess of non-radiolabeled Example-1 (2 μM). Specific binding was calculated by subtracting the non-specific signal from the total signal. Competition was calculated as a percentage, where 0% was defined as specific binding in the presence of the solvent, and 100% was defined as the value obtained in the presence of an excess of non-radiolabeled Example-1. Ki values ​​were calculated in GraphPad Prism7 using a unit-point specific binding model with nonlinear regression curve fitting. All measurements were performed at least twice technically. For compounds tested in more than one experiment, the mean or Ki value of the replicates from the independent experiments was reported.

[1255] result Evaluation of the compounds in Example 1 and Example-1 3 [H-1] ligand competitive binding potency to α-syn aggregates derived from the brains of PD patients. Results of the microradioactive binding competition assays of the tested compounds are shown in Table 8: competition percentages at 1 μM and 100 nM. Table 8 also shows K... i value.

[1256] Table 8

[1257]

[1258]

[1259]

[1260]

[1261]

[1262]

[1263] Table 8: Evaluation of binding affinity of human PD brain-derived α-syn aggregates by microradioactive binding competition assay. Relative to tritized Example-1 in the presence of Compounds 2 to 181 at 1 μM and 100 nM. 3The percentage (%) of competition for the H-1] ligand is also shown. Ki values ​​for the selected example compounds are also displayed. * Average Ki values ​​from independent experiments using homogenates of PD-derived brain from three different donors. ** Average Ki values ​​from independent experiments using homogenates of PD-derived brain from two different donors. As shown in Table 8, example compounds 2-181 of the present invention exhibit efficient binding to PD-derived α-syn aggregates.

[1264] 11. Example 4 [ 3 Evaluation of target binding of H-4 in α-synucleinopathies

[1265] 11A: High-resolution autoradiography

[1266] This scheme is adapted from Marquie et al., 2015. The slices were compared with compound 4 of Example 4 (Example-4 [...]). 3 H-4]) or reference Tau ligand ([ 3 Incubate [H]-Tau-Ref, 20 nM) at RT for 1 hour. Then wash the sections as follows: wash once in ice-cold 50 mM Tris-HCl pH 7.4 buffer for 1 minute each time, wash twice in ice-cold 70% ethanol for 1 minute each time, wash once in ice-cold 50 mM Tris-HCl pH 7.4 buffer, and finally rinse briefly in ice-cold distilled water. The sections were then dried and exposed to Ilford Nuclear Emulsion Type K5 (AgarScientific, AGP9281) in a light-shielded slide storage box. Five days later, the sections were developed by immersing them sequentially in the following solutions: 1.) Ilford Phennesol developer (1:5 diluted in H2O, Agar Scientific, AGP 9106), 2.) IlfoStop solution (1:20 diluted in H2O, Agar Scientific, AGP 9104), 3.) Ilford Hypam fixer (1:5 diluted in H2O, Agar Scientific, AGP 9183), and finally rinsed with H2O.

[1267] Immunostaining was also performed on the same sections during display. For image acquisition, sections were fixed using ProLong Gold anti-fading reagent (Invitrogen P36930) and imaged on a Panoramic 150 slide scanner (3DHistech) with 20x objectives, capturing bright-field and fluorescence images, respectively.

[1268] 11B. By using antibody-stained sections

[1269] Immunostaining of brain sections was performed using a commercially available antibody specific to phosphorylated serine α-synuclein at amino acid 129 (α-syn-pS129, rabbit monoclonal antibody, Abcam 51253). Sections were fixed with 4% formaldehyde (Sigma, 252549) at 4°C for 15 minutes and washed three times with 1×PBS (Dulbecco phosphate-buffered saline, Sigma D1408) at room temperature for 5 minutes each time. Next, sections were saturated with blocking buffer (PBS, 10% NGS, 0.25% Triton X-100) at RT for 1 hour and incubated overnight at 4°C with a primary antibody corresponding to α-syn-pS129. The next day, sections were washed three times with 1×PBS for 5 minutes each time and then incubated with Alexa Fluor 647-labeled goat anti-human IgG secondary antibody. Rabbit (Abcam, ab150079) antibody was incubated at room temperature for 45 minutes. After incubation with secondary antibody, the slides were washed three times in PBS and then processed further. For image acquisition, the slides were fixed with ProLong Gold anti-fading reagent (Invitrogen P36930) and imaged using a Panoramic 150 slide scanner (3DHistech; Hungary).

[1270] result Example 4 3 [H-4] High-resolution autoradiography was performed on frozen human brain slices from PD donors. Accumulated silver particles were detected from samples from Example-4 [ 3 The strong autoradiographic signal of H-4] Figure 12 (Lower part), and co-localized with the immunofluorescence signal from the α-syn-pS129 antibody ( Figure 12 (Upper part) indicates strong target binding on Lewy bodies and Lewy neurites, as well as on very small α-syn aggregates in PD tissue.

[1271] 12. Autoradiography of brain slices from PD, MSA and non-dementia control (NDC) donors - Example 4 [ 3 Evaluation of specific binding to [H-4]

[1272] First, frozen human brain sections from one case of familial PD (α-synuclein [SNCA] gene G51D missense mutation) (labeled SNCA), one case of idiopathic PD, one case of MSA, and two non-dementia controls (NDC) were briefly fixed at 4°C with 4% paraformaldehyde (Sigma, 252549) for 15 minutes and washed three times with PBS (Dulbecco phosphate-buffered saline, Sigma) for 5 minutes each at room temperature. Then, all slides were equilibrated in 50 mM Tris-HCl pH 7.4 buffer for 20 minutes before use in experiments. Each brain section was then mixed with a fixed concentration (10 nM) of tritium-modified compound 4 (Example-4) in Tris-HCl buffer. 3 H-4]) was incubated at room temperature for 2 hours (total binding, 'total'). To determine nonspecific binding, Example-4 [ 3 [H-4] was mixed with 5 mM of a non-radioactively labeled compound (Example 4, 'NSB'). The slide was washed and then exposed and scanned in a real-time autoradiography system (Beaquant Instrument, ai4R).

[1273] result: Example 4 3 [H-4] showed specific binding in various α-synucleinopathy tissues (including MSA cases, familial PD cases, and idiopathic PD cases). Figure 13A Compared to non-dementia controls, diseased donors showed stronger autoradiographic signals, confirming target binding and exhibiting a strong correlation with the distribution of pathological α-synuclein load. Figure 13B Additionally, Example 4 [ 3 H-4] showed a replaceable signal in various α-synucleinosis cases examined and a very weak signal in several non-disease control cases.

[1274] 13. Saturation binding study of α-syn aggregates from PD-derived brain using microradiotherapy

[1275] PD-derived α-syn aggregates were spotted onto a microarray slide. The slide was then subjected to increasing concentrations in the range of 1.56 nM to 80 nM, as described in Example 4. 3H-4 was incubated together. After incubation, the slide was scanned using a real-time autoradiography system (Beaquant Instrument, AI4R). The signal was quantified using Beamage image analysis software (ai4R). Non-specific signals were measured using an excess of non-radioactively labeled reference ligand (Example-4, 2 μM), and specific binding was calculated by subtracting the non-specific signal from the total signal. K was calculated in GraphPad Prism7 using a unit-point specific binding model with nonlinear regression curve fitting. d value.

[1276] result Example 4 was evaluated in a study of the saturation binding of homogenates in PD tissues via microradiation. 3 H-4]( Figure 14 ). Calculate the dissociation constant (K) at 21 nM. d ()( Figure 14 (Table 9) shows a good binding affinity for pathological α-synuclein aggregates.

[1277] Table 9:

[1278] <![CDATA[Example - 4 3 H - 4]]]> PD homogenate Kd 21nM <![CDATA[R 2 ]]> 0.86

[1279] Table 9: Effects of microradiation on Example-4 [ 3 [H-4] Assessment of the binding affinity of homogenates from human PD brain tissue. The dissociation constant (Kd) and binding site occupancy (B) were calculated using a single-point specific binding model in GraphPad Prism 7 and by applying nonlinear regression curve fitting. max R 2 It is the coefficient of determination.

[1280] 14. Used to determine the inhibitory constant (K) of compound 4 in Example 4 on AD brain homogenate. i Radioactive binding competitive assay

[1281] Human Alzheimer's disease (AD) brain homogenate was prepared according to the method disclosed in Example 7 (see above).

[1282] A fixed concentration of the AD insoluble fraction was compared with a 10 nM tritized reference Aβ ligand ([ 3Compound 1 of Example, containing non-radiolabeled reference ligands in the concentration range of 400 pM to 2 μM (H-Aβ-Ref), was incubated at RT for 2 h. The sample was then vacuum filtered in a GF / C filter plate (PerkinElmer) to capture aggregates with bound radioligands and washed five times with 50 mM Tris pH 7.5. The GF / C filter was then dried and scintillation solution (Ultimate Gold, PerkinElmer) was added to each well. The filter was analyzed on a MicroBeta2 scintillation counter (PerkinElmer). Non-specific signals were determined with an excess of the non-radiolabeled reference ligand (2 μM), and specific binding was calculated by subtracting the non-specific signal from the total signal. Competition was calculated as a percentage, where 0% was defined as specific binding in the presence of the solvent and 100% as the value obtained in the presence of an excess of the non-radiolabeled reference ligand. Ki values ​​were calculated in GraphPad Prism 7 by applying a nonlinear regression curve fit using a single-point specific binding model. Measurements were performed in two independent experiments with two technical replicates.

[1283] result :like Figure 15 As shown in Table 10, the Ki value of Compound 4 from Example 4 was determined at 297 nM in the AD brain-derived homogenate. Based on the binding of Compound 4 by microradiography... 3 The binding affinity of H-4] in PD brain homogenates, as reported in Example 13 (above) with a value of 21 nM, and its specific binding in α-synucleinopathy brain tissue by autoradiography, showed that Compound 4 of Example 4 exhibited good selectivity for α-synuclein compared to Aβ pathological aggregates present in human brain homogenates. Furthermore, compared to the reference Tau binder used as a positive control, Example-4 [ 3 H-4] did not show specific target binding on Tau aggregates in AD brain tissue. Figure 16 The results indicate good selectivity compared to Tau pathological aggregates. In summary, these data demonstrate that Compound 4 of Examples exhibits the desired selectivity for α-syn aggregates.

[1284] Table 10:

[1285]

[1286] Table 10: Ki value determination of Compound 4 of Example 4 by replacing [3H]-Aβ-Ref with non-radioactively labeled Compound 4 on AD brain-derived homogenates. Ki and R² values ​​were calculated by applying nonlinear regression curve fitting using a single-point specific binding model of GraphPad Prism7.

[1287] 15. First-in-human (FIH) study

[1288] Conduct an assessment 18 Example F-1, as a phase 1 study of potential PET radioligands, was used to image α-synuclein deposits in the brains of patients with suspected α-synuclein pathology, compared to healthy volunteers. The aim of this study was to characterize individuals with suspected idiopathic Parkinson's disease (PD) and healthy volunteers (HV). 18 F - Safety, imaging, and pharmacokinetic characteristics of Example 1. A total of up to 10 subjects can be recruited (targeting up to 5 HV subjects and up to 5 subjects with idiopathic PD).

[1289] Selection criteria for all individuals:

[1290] • Individuals must be able to provide written informed consent, which must be obtained before any evaluation is conducted.

[1291] • Female individuals must be fertile, or if they are fertile, they must consent to the use of contraception and not to donate eggs. Individuals without a record of being fertile, as determined by the researchers, can undergo pregnancy testing.

[1292] • Male individuals of reproductive potential must commit to using two methods of contraception, one of which is a barrier method used during the study and for 90 days after the study ends.

[1293] Male individuals are prohibited from donating sperm during the study period and for 90 days after the study is completed.

[1294] • For subjects undergoing arterial cannulation, adequate hand circulation is used for safe placement of the arterial line (as determined by the Allen test) and coagulation (prothrombin time [PT] and partial prothrombin time [PTT]).

[1295] • If an individual is taking bupropion, the individual must agree to keep the medication for at least 12 hours before DaTscan imaging (if performed).

[1296] Additional inclusion criteria for HV individuals:

[1297] • Men and women aged 21 and above.

[1298] • Healthy, with no clinically relevant findings during physical examinations at screening and when reporting tracer imaging visits to clinics.

[1299] • No family history of alpha-synucleinopathies (including PD) or other early-onset neurological disorders associated with dementia.

[1300] • No clinically significant personal history of neurological and / or mental disorders.

[1301] • No evidence of dopamine transporter deficiency on a dopamine active transporter (DaT) scan performed as part of the screening or on a previously obtained DaTscan (within 6 months prior to signing the consent form).

[1302] • Has a Montreal Cognitive Assessment (MoCA) score of ≥26.

[1303] • For example, no cognitive impairment as determined by the supervisor (PI).

[1304] Additional selection criteria for subjects with α-synucleinopathy:

[1305] • Men and women aged 40 and above.

[1306] • Individuals diagnosed with any of the following:

[1307] о Idiopathic PD

[1308] оPD with genetic risk factors (excluding leucine-rich repeat unit kinase 2 [LRRK2] mutations)

[1309] • Brain magnetic resonance imaging (MRI) consistent with the diagnosis of alpha-synucleinopathy showed no evidence of focal disease to explain individual neurological symptoms.

[1310] • Evidence of dopamine transporter defects on DaTscan as part of a screening process or on previously obtained DaTscanscans.

[1311] • Medications used to treat the symptoms of alpha-synucleinosis must maintain a stable dosage regimen for at least 30 days prior to the screening visit.

[1312] • Able to tolerate lying in the scanner for up to approximately 180 minutes without excessive head or jaw tremors or movement disorders sufficient to cause significant motion artifacts on a PET scan.

[1313] After recruitment, subjects received one intravenous injection. 18 Example F - No more than 10 mCi. Visual inspection and quantitative assessment. 18 Example F - Brain uptake and pharmacokinetics in human individuals, and obtaining safety data. This included suspected idiopathic PD cases. 18 F-Comparison of PET signals with HV cross-sections in Example 1.

[1314] 16: Formulation

[1315] 18 F. Capture and Elution: [ 18The F-fluoride was transferred to and captured on an ion exchange column. It was then eluted into the reaction vessel (RV1) with an aqueous solution of potassium carbonate (1.6 mg) and Kryptofix 222 (10 mg) in acetonitrile. The solution was first evaporated by heating at 95 °C for 4 min under vacuum and helium. Then, acetonitrile (1 mL) was added to RV1, and evaporation continued for 2 min under the same conditions under vacuum and helium. After a second addition of acetonitrile (1 mL), a final evaporation was carried out at 95 °C for 2 min under vacuum and helium. Finally, the reactor was cooled to 60 °C.

[1316] Radiolabeling reaction: A solution of the precursor (1.0 mg) in anhydrous dimethyl sulfoxide was added to the reaction vessel, and the reaction mixture was heated at 100 °C for 10 min. The reactor was cooled to 40 °C, diluted with HPLC mobile phase (1.8 mL), and the contents were transferred to a loop loading vial (RV2). The reactor was rinsed with water for injection (2.5 mL), and the rinse solution was transferred to RV2. The contents of RV2 were transferred to an HPLC syringe loop for purification.

[1317] Purification and formulation of the pharmaceutical product: Purification was performed by HPLC using a semi-preparative Agilent Eclipse XDB C18 column (5 μm, 250 x 9.4 mm) and eluted with a methanol / ammonium acetate mixture (20 mM, 50 / 50, v / v) at a flow rate of 4 mL / min. The product fraction was collected in a flask containing 20 mL of sodium ascorbate (5 mg / mL) in water for injection (WFI). The diluted product mixture was passed through a C18 solid-phase extraction column and washed with 10 mL of sodium ascorbate (5 mg / mL) in WFI. The radiolabeled product was eluted from the SPE column to a formulation flask pre-filled with 10 mL of sodium ascorbate (10 mg / mL) in saline with 1.0 mL of 200 standard strength USP grade ethanol. The column was washed with 4.0 mL of sodium ascorbate in saline (10 mg / mL) and the wash solution was mixed with the contents of the formulation flask. The resulting solution was passed through a sterile 0.2 μm membrane filter into a sterile filter connecting vial (final product vial, FPV) pre-filled with 15 mL of physiological saline.

[1318] The stability of the radiolabeled product over time was studied, and it was verified that it remained within specifications for 8 hours after synthesis.

[1319] The batch formulation quantities are shown in Table 11:

[1320] Precursor <![CDATA[1mg a ]]> [18F] Fluoride <4Ci physiological saline 50mL ethanol 1mL Sodium ascorbate 500mg

[1321] a Removed during processing

[1322] The final formulation of the radiolabeled product developed for this study has a volume of 30 mL, and the aim is to achieve the following content based on an injection volume of 10 mL in the final dosage form, as shown in Table 12:

[1323] Amount of radioactivity carrier physiological saline Sodium ascorbate ethanol ≤10mCi ≤10μg ≤9.67ml ≤46.7mg ≤0.33ml

Claims

1. Compounds of formula (I) (I) or its use 3 H or 18 F-labeled compounds, stereoisomers, racemic mixtures, and pharmaceutically acceptable salts. in As a heteroaryl group, it is selectively chosen from the following: 、 、 and , R 0 It is H or C1-C4 alkyl; R 1 -CN; or halogen; or C1-C4 alkyl; or C1-C4 alkoxy; or -N(C1-C4 alkyl)2; or -NH(C1-C4 alkyl); or H; or R 1 It is a -NH-C3-C6 cycloalkyl, C3-C6 cycloalkyl or a 4- to 6-membered heterocyclic group containing one or two heteroatoms selected from N, O or S, each of which is optionally substituted with at least one halogen; R 2 It is an aryl, 5-membered, or 6-membered heteroaryl group, wherein R 2 Selected from the following: , in R 2a R 2a’ Independently selected from H or F; R 2b It is independently selected from F, OH, C1-C4 alkyl, halo-C1-C4 alkyl, NH2, CN or C1-C4 alkoxy; R 2c R 2c’ It is independently selected from H, F, OH, OCH3 or CH3; R 2d Selected from H, F, or OH; R 2e Selected from H, OH, CH3 or F; Z can be N, NH, N (C1-C4 alkyl), N (halogenated C1-C4 alkyl), O, or S independently; Z 1 It can be N, NH, O, or S independently; p is 0, 1, or 2; m is 0 or 1; When valence is allowed For combinations of single and double bonds; and This indicates the location of the bond.

2. The compound of claim 1, having formula (IIa) or (IIb), (IIa) or (IIb), or its use 3 H or 18 F-labeled compounds, stereoisomers, racemic mixtures, and pharmaceutically acceptable salts.

3. The compound of claim 1, having formula (IIIa) or (IIIb), (IIIa) or (IIIb) or its use 3 H or 18 F-labeled compounds, stereoisomers, racemic mixtures, and pharmaceutically acceptable salts.

4. The compound of claim 1 or 3, wherein R 1 It is a -NH-C3-C6 cycloalkyl, C3-C6 cycloalkyl or a 4- to 6-membered heterocyclic group containing one or two heteroatoms selected from N, O or S, each of which is optionally substituted with at least one halogen.

5. The compound of claim 4, wherein R 1 Selected from the following: 。 6. A compound, wherein the compound is selected from the group consisting of, or thereof, compounds selected from, or thereof. 3 H or 18 F-labeled compounds, stereoisomers, racemic mixtures, and pharmaceutically acceptable salts: 。 7. A compound of formula (I), wherein the compound is used... 3 H or 18 F-labeled compounds, and said compounds as defined in any one of claims 1-6.

8. The compound of formula (I) of claim 7, wherein R 1 for , , or .

9. The compound of claim 7, wherein the use of 3 H or 18 F-labeled compounds are , Or its pharmaceutically acceptable salt.

10. Use of the compound of any one of claims 7-9 or its stereoisomers, racemic mixtures, or pharmaceutically acceptable salts for the preparation of a diagnostic imaging agent for imaging α-synuclein aggregates, said α-synuclein aggregates including, but not limited to, Lewy bodies and / or Lewy neurites.

11. Use of the compound of any one of claims 7-9 or its stereoisomers, racemic mixtures, or pharmaceutically acceptable salts for the preparation of a diagnostic imaging agent for imaging α-synuclein aggregates by positron emission tomography, said α-synuclein aggregates including, but not limited to, Lewy bodies and / or Lewy neurites.

12. The use according to claim 10, wherein the use is for in vitro imaging, ex vivo imaging or in vivo imaging.

13. The use according to claim 12, wherein the use is for in vivo imaging.

14. The use according to claim 12, wherein the use is for brain imaging.

15. The use according to claim 14, wherein the diagnostic imaging agent is used for the diagnosis of diseases, conditions, or abnormalities or susceptibility to α-synuclein aggregates, including but not limited to Lewy bodies and / or Lewy neurites, wherein the diseases, conditions, or abnormalities are optionally selected from Parkinson's disease, SNCA repeat carriers, Lewy body dementia, Parkinson's disease dementia, diffuse Lewy body disease, Alzheimer's disease, Down syndrome, multiple system atrophy, traumatic brain injury, and chronic injury. Encephalopathy, boxer's dementia, tau disease, Creutzfeldt-Jakob disease, Huntington's disease, motor neuron disease, amyotrophic lateral sclerosis, neurodegeneration with type 1 brain iron accumulation, prions, ataxia-telangiectasia, idiopathic orofacial motor disorder, subacute sclerosing panencephalitis, Gerstmann-Straussler-Scheinker syndrome, inclusion body myositis, Gaucher disease, Krabby disease, and other lysosomal storage diseases and oculomotor sleep behavior disorders.

16. The use according to claim 15, wherein the Alzheimer's disease is selected from sporadic Alzheimer's disease, familial Alzheimer's disease with APP mutation, familial Alzheimer's disease with PS-1, PS-2 or other mutations, familial British dementia, Lewy body variant of Alzheimer's disease, and Lewy body dementia is Lewy body dementia.

17. The use according to claim 15, wherein Parkinson's disease includes sporadic, familial, and non-familial types of Parkinson's disease with mutations other than α-synuclein, simple autonomic failure, and Lewy body dysphagia; Lewy body dementia includes "simple" Lewy body dementia; multiple system atrophy includes Shy-Drager syndrome, striatal substantia nigra degeneration, and olivary pontocerebellar atrophy; tau disorders include Pick's disease, frontotemporal dementia, progressive supranuclear palsy, corticobasal degeneration, and Niemann-Pick type C1 disease, and frontotemporal dementia associated with Parkinson's syndrome related to chromosome 17; amyotrophic lateral sclerosis includes sporadic, familial, and Guam ALS-dementia syndrome; neurodegeneration with type 1 brain iron accumulation includes Hastings-Schwarz syndrome; and other lysosomal storage disorders include Kufor-Rakeb syndrome and Sanfilippo syndrome.

18. The use according to claim 15, wherein the disease is Parkinson's disease.

19. The use according to claim 15, wherein the disease is multiple system atrophy.

20. The use according to claim 15, wherein the disease is Lewy body dementia.

21. The use according to claim 15, wherein the disease is Parkinson's disease or dementia.

22. The use according to claim 15, wherein the disease is an SNCA repeat carrier.

23. The use according to claim 15, wherein the disease is Alzheimer's disease.

24. The use according to any one of claims 10-23, wherein the use is for a person.

25. A diagnostic composition comprising the compound of any one of claims 7-9 or a stereoisomer thereof, a racemic mixture, a pharmaceutically acceptable salt, and at least one pharmaceutically acceptable excipient, carrier, or adjuvant.

26. The diagnostic composition of claim 25, wherein the excipient is a diluent.

27. Use of the compound of any one of claims 7-9 or its stereoisomers, racemic mixtures, pharmaceutically acceptable salts, or the diagnostic composition of claim 25 for the preparation of a diagnostic imaging agent for imaging diseases, conditions, or abnormalities in an individual associated with α-synuclein aggregates, including but not limited to Lewy bodies and / or Lewy neurites.

28. Use of the compound of any one of claims 7-9 or its stereoisomers, racemic mixtures, pharmaceutically acceptable salts, or the diagnostic composition of claim 25 for the preparation of a diagnostic imaging agent for positron emission tomography imaging of α-synuclein aggregates in individual tissues, including but not limited to Lewy bodies and / or Lewy neurites. The tissues mentioned are central nervous system tissues, eye tissues, or brain tissues.

29. The use according to claim 28, wherein the tissue is brain tissue.

30. Use of the compound of any one of claims 7-9 or its stereoisomers, racemic mixtures, pharmaceutically acceptable salts, or the diagnostic composition of claim 25 for the preparation of a diagnostic imaging agent for the detection of neurological diseases, conditions, or abnormalities associated with α-synuclein aggregates in an individual, including but not limited to Lewy bodies and / or Lewy neurites.

31. Use of the compound of any one of claims 7-9 or its stereoisomers, racemic mixtures, pharmaceutically acceptable salts, or the diagnostic composition of claim 25 for the preparation of a diagnostic imaging agent for the detection and / or quantification of α-synuclein aggregates in individual tissues, including but not limited to Lewy bodies and / or Lewy neurites.

32. Use of the compound of any one of claims 7-9 or its stereoisomers, racemic mixtures, pharmaceutically acceptable salts, or the diagnostic composition of claim 25 for the preparation of a diagnostic imaging agent for the diagnostic imaging of an individual's brain.

33. Use of the compound of any one of claims 7-9 or its stereoisomers, racemic mixtures, pharmaceutically acceptable salts, or the diagnostic composition of claim 25 for the preparation of a diagnostic imaging agent for the purpose of diagnosing diseases, conditions, or abnormalities associated with α-synuclein aggregates, including but not limited to Lewy bodies and / or Lewy neurites.

34. The use of the compound of any one of claims 7-9 or its stereoisomers, racemic mixtures, pharmaceutically acceptable salts, or the diagnostic composition of claim 25 for the preparation of a diagnostic imaging agent for collecting data on susceptibility to diseases, conditions, or abnormalities associated with α-synuclein aggregates, including but not limited to Lewy bodies and / or Lewy neurites.

35. The use of the compound of any one of claims 7-9 or its stereoisomers, racemic mixtures, pharmaceutically acceptable salts, or the diagnostic composition of claim 25 for the preparation of a diagnostic imaging agent for acquiring data for predicting the prognosis of diseases, conditions, or abnormalities associated with α-synuclein aggregates, including but not limited to Lewy bodies and / or Lewy neurites.

36. Use of the compound of any one of claims 7-9 or its stereoisomers, racemic mixtures, pharmaceutically acceptable salts, or the diagnostic composition of claim 25 for the preparation of a diagnostic imaging agent for collecting data on diseases, conditions, or abnormalities associated with α-synuclein aggregates in patients, including but not limited to Lewy bodies and / or Lewy neurites.

37. Use of the compound of any one of claims 7-9 or its stereoisomers, racemic mixtures, pharmaceutically acceptable salts, or the diagnostic composition of claim 25 for the preparation of a diagnostic imaging agent for collecting data on the responsiveness of patients with diseases, conditions, or abnormalities associated with α-synuclein aggregates to treatments for such diseases, conditions, or abnormalities, including but not limited to Lewy bodies and / or Lewy neurites.

38. Compounds of formula (IV-F) (IV-F) Or its stereoisomers, racemic mixtures, or pharmaceutically acceptable salts, wherein R 3 Selected from , , or ; R 4 It is an aryl or 5- or 6-membered heteroaryl, wherein R 4 Selected from: ， in R 2a R 2a’ Independently selected from H or F; R 2b Independently selected from F, -OH, C1-C4 alkyl, halo-C1-C4 alkyl, -NH2, -CN or C1-C4 alkoxy; R 2c R 2c’ It is independently selected from H, F, OH, OCH3 or CH3; R 2d Selected from H, F, or -OH; R 2e Selected from H, OH, CH3 or F; Z can be N, NH, N (C1-C4 alkyl), N (halogenated C1-C4 alkyl), O, or S independently; Z 1 It can be N, NH, O, or S independently; p is 0, 1, or 2; m is 0 or 1; When valence is allowed It is a combination of single and double bonds; and The location of the bond. LG is selected from halogen, C 1–4 Alkyl sulfonates and C 6–10 Aryl sulfonates, wherein C 6–10 The aryl group can be optionally replaced by –CH3 or –NO2.

39. The compound of formula (IV-F) of claim 38, wherein LG is selected from bromine, chlorine, iodine, C 1–4 Alkyl sulfonates and C 6–10 Aryl sulfonates, wherein C 6–10 The aryl group can be optionally replaced by –CH3 or –NO2.

40. The compound of formula (IV-F) of claim 38 or 39, wherein it is ； Or a pharmaceutically acceptable salt thereof, wherein LG is a methanesulfonate or a nitrobenzenesulfonate.

41. Compounds of formula (IV-H) (IV-H) Or its stereoisomers, racemic mixtures, or pharmaceutically acceptable salts, wherein R 5 Selected from , , , , , , , or ; R 6 It is an aryl or 5- or 6-membered heteroaryl, wherein R 6 Selected from the following: in R 2a R 2a’ Independently selected from H, X, or F; R 2b Independently selected from X, F, -OH, C1-C4 alkyl, halo-C1-C4 alkyl, -NH2, -CN or C1-C4 alkoxy, wherein the C1-C4 alkyl, halo-C1-C4 alkyl or C1-C4 alkoxy optionally includes one or more X; R 2c R 2c’ It is independently selected from X, H, F, OH, OCH3 or CH3; R 2d Selected from X, H, F or -OH; R 2e Selected from X, H, OH, CH3, or F; Z can be N, NH, N (C1-C4 alkyl), N (halogenated C1-C4 alkyl), O, or S independently; Z 1 It can be N, NH, O, or S independently; p is 0, 1, or 2; m is 0 or 1; When valence is allowed It is a combination of single and double bonds; The location of the bond; Fluorine is 19 F; X is bromine, chlorine, or iodine; and Where R 6 It contains at least one X.

42. The compound of formula (IV-H) according to claim 41, wherein: , Or its pharmaceutically acceptable salt.

43. Compounds of formula (IV-J) (IV-J), Or its stereoisomers, racemic mixtures, or pharmaceutically acceptable salts, wherein R 7 Selected from , , , , , , , or ; R 8 Selected from the following: in R 2a R 2a’ Independently selected from H or F; R 2b Independently selected from F, -OH, C1-C4 alkyl, halo-C1-C4 alkyl, -NH2, -CN or C1-C4 alkoxy; p is 0, 1, or 2; R z Selected from H, C1-C4 alkyl, and halo-C1-C4 alkyl; When valence is allowed It is a combination of single and double bonds; Fluorine is 19 F; and This indicates the location of the bond.

44. The compound of formula (IV-J) according to claim 43, wherein: , Or its pharmaceutically acceptable salt.

45. A method for preparing the compound of claim 7, 8 or 9, comprising reacting the compound of any one of claims 38-40 with... 18 The F-fluorinating agent reaction causes LG to be... 18 F substitution.

46. ​​The method of claim 45, wherein 18 F-fluorinating agents are selected from K 18 F, Rb 18 F, Cs 18 F, Na 18 F, Rb 18 F, Kryptofix[222]K 18 F, 18 F's four (C) 1-6 alkyl)ammonium salts and [ 18 F]Tetrabutylammonium fluoride.

47. A method for preparing the compound of claim 7, 8 or 9, comprising mixing the compound of claim 41 or 42 with... 3 H radiolabeled reagent reaction.

48. A method for preparing the compound of claim 7, 8, or 9, comprising reacting the compound of claim 40 or 41 with a CT3 radiolabeled reagent, wherein T is... 3 H.

49. A kit for detecting and / or diagnosing diseases, conditions or abnormalities associated with α-synuclein aggregates, wherein the kit comprises at least one compound of formula (I) as defined in any one of claims 1-9, or a stereoisomer thereof, a racemic mixture, or a pharmaceutically acceptable salt thereof.

50. A kit for preparing radiopharmaceutical formulations, wherein the kit comprises a sealed vial containing at least one compound as defined in any one of claims 38-44.

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